| anno_start anno_end anno_text entity_type sentence section | |
| 61 70 Regnase-1 protein Structural basis for the regulation of enzymatic activity of Regnase-1 by domain-domain interactions TITLE | |
| 0 9 Regnase-1 protein Regnase-1 is an RNase that directly cleaves mRNAs of inflammatory genes such as IL-6 and IL-12p40, and negatively regulates cellular inflammatory responses. ABSTRACT | |
| 16 21 RNase protein_type Regnase-1 is an RNase that directly cleaves mRNAs of inflammatory genes such as IL-6 and IL-12p40, and negatively regulates cellular inflammatory responses. ABSTRACT | |
| 44 49 mRNAs chemical Regnase-1 is an RNase that directly cleaves mRNAs of inflammatory genes such as IL-6 and IL-12p40, and negatively regulates cellular inflammatory responses. ABSTRACT | |
| 80 84 IL-6 protein_type Regnase-1 is an RNase that directly cleaves mRNAs of inflammatory genes such as IL-6 and IL-12p40, and negatively regulates cellular inflammatory responses. ABSTRACT | |
| 89 97 IL-12p40 protein_type Regnase-1 is an RNase that directly cleaves mRNAs of inflammatory genes such as IL-6 and IL-12p40, and negatively regulates cellular inflammatory responses. ABSTRACT | |
| 20 30 structures evidence Here, we report the structures of four domains of Regnase-1 from Mus musculus—the N-terminal domain (NTD), PilT N-terminus like (PIN) domain, zinc finger (ZF) domain and C-terminal domain (CTD). ABSTRACT | |
| 50 59 Regnase-1 protein Here, we report the structures of four domains of Regnase-1 from Mus musculus—the N-terminal domain (NTD), PilT N-terminus like (PIN) domain, zinc finger (ZF) domain and C-terminal domain (CTD). ABSTRACT | |
| 65 77 Mus musculus species Here, we report the structures of four domains of Regnase-1 from Mus musculus—the N-terminal domain (NTD), PilT N-terminus like (PIN) domain, zinc finger (ZF) domain and C-terminal domain (CTD). ABSTRACT | |
| 82 99 N-terminal domain structure_element Here, we report the structures of four domains of Regnase-1 from Mus musculus—the N-terminal domain (NTD), PilT N-terminus like (PIN) domain, zinc finger (ZF) domain and C-terminal domain (CTD). ABSTRACT | |
| 101 104 NTD structure_element Here, we report the structures of four domains of Regnase-1 from Mus musculus—the N-terminal domain (NTD), PilT N-terminus like (PIN) domain, zinc finger (ZF) domain and C-terminal domain (CTD). ABSTRACT | |
| 107 127 PilT N-terminus like structure_element Here, we report the structures of four domains of Regnase-1 from Mus musculus—the N-terminal domain (NTD), PilT N-terminus like (PIN) domain, zinc finger (ZF) domain and C-terminal domain (CTD). ABSTRACT | |
| 129 132 PIN structure_element Here, we report the structures of four domains of Regnase-1 from Mus musculus—the N-terminal domain (NTD), PilT N-terminus like (PIN) domain, zinc finger (ZF) domain and C-terminal domain (CTD). ABSTRACT | |
| 142 153 zinc finger structure_element Here, we report the structures of four domains of Regnase-1 from Mus musculus—the N-terminal domain (NTD), PilT N-terminus like (PIN) domain, zinc finger (ZF) domain and C-terminal domain (CTD). ABSTRACT | |
| 155 157 ZF structure_element Here, we report the structures of four domains of Regnase-1 from Mus musculus—the N-terminal domain (NTD), PilT N-terminus like (PIN) domain, zinc finger (ZF) domain and C-terminal domain (CTD). ABSTRACT | |
| 170 187 C-terminal domain structure_element Here, we report the structures of four domains of Regnase-1 from Mus musculus—the N-terminal domain (NTD), PilT N-terminus like (PIN) domain, zinc finger (ZF) domain and C-terminal domain (CTD). ABSTRACT | |
| 189 192 CTD structure_element Here, we report the structures of four domains of Regnase-1 from Mus musculus—the N-terminal domain (NTD), PilT N-terminus like (PIN) domain, zinc finger (ZF) domain and C-terminal domain (CTD). ABSTRACT | |
| 4 7 PIN structure_element The PIN domain harbors the RNase catalytic center; however, it is insufficient for enzymatic activity. ABSTRACT | |
| 27 32 RNase protein_type The PIN domain harbors the RNase catalytic center; however, it is insufficient for enzymatic activity. ABSTRACT | |
| 33 49 catalytic center site The PIN domain harbors the RNase catalytic center; however, it is insufficient for enzymatic activity. ABSTRACT | |
| 18 21 NTD structure_element We found that the NTD associates with the PIN domain and significantly enhances its RNase activity. ABSTRACT | |
| 42 45 PIN structure_element We found that the NTD associates with the PIN domain and significantly enhances its RNase activity. ABSTRACT | |
| 84 89 RNase protein_type We found that the NTD associates with the PIN domain and significantly enhances its RNase activity. ABSTRACT | |
| 4 7 PIN structure_element The PIN domain forms a head-to-tail oligomer and the dimer interface overlaps with the NTD binding site. ABSTRACT | |
| 23 35 head-to-tail protein_state The PIN domain forms a head-to-tail oligomer and the dimer interface overlaps with the NTD binding site. ABSTRACT | |
| 36 44 oligomer oligomeric_state The PIN domain forms a head-to-tail oligomer and the dimer interface overlaps with the NTD binding site. ABSTRACT | |
| 53 68 dimer interface site The PIN domain forms a head-to-tail oligomer and the dimer interface overlaps with the NTD binding site. ABSTRACT | |
| 87 103 NTD binding site site The PIN domain forms a head-to-tail oligomer and the dimer interface overlaps with the NTD binding site. ABSTRACT | |
| 15 24 mutations experimental_method Interestingly, mutations blocking PIN oligomerization had no RNase activity, indicating that both oligomerization and NTD binding are crucial for RNase activity in vitro. ABSTRACT | |
| 34 37 PIN structure_element Interestingly, mutations blocking PIN oligomerization had no RNase activity, indicating that both oligomerization and NTD binding are crucial for RNase activity in vitro. ABSTRACT | |
| 61 66 RNase protein_type Interestingly, mutations blocking PIN oligomerization had no RNase activity, indicating that both oligomerization and NTD binding are crucial for RNase activity in vitro. ABSTRACT | |
| 118 121 NTD structure_element Interestingly, mutations blocking PIN oligomerization had no RNase activity, indicating that both oligomerization and NTD binding are crucial for RNase activity in vitro. ABSTRACT | |
| 146 151 RNase protein_type Interestingly, mutations blocking PIN oligomerization had no RNase activity, indicating that both oligomerization and NTD binding are crucial for RNase activity in vitro. ABSTRACT | |
| 27 36 Regnase-1 protein These results suggest that Regnase-1 RNase activity is tightly controlled by both intramolecular (NTD-PIN) and intermolecular (PIN-PIN) interactions. ABSTRACT | |
| 37 42 RNase protein_type These results suggest that Regnase-1 RNase activity is tightly controlled by both intramolecular (NTD-PIN) and intermolecular (PIN-PIN) interactions. ABSTRACT | |
| 98 101 NTD structure_element These results suggest that Regnase-1 RNase activity is tightly controlled by both intramolecular (NTD-PIN) and intermolecular (PIN-PIN) interactions. ABSTRACT | |
| 102 105 PIN structure_element These results suggest that Regnase-1 RNase activity is tightly controlled by both intramolecular (NTD-PIN) and intermolecular (PIN-PIN) interactions. ABSTRACT | |
| 127 130 PIN structure_element These results suggest that Regnase-1 RNase activity is tightly controlled by both intramolecular (NTD-PIN) and intermolecular (PIN-PIN) interactions. ABSTRACT | |
| 131 134 PIN structure_element These results suggest that Regnase-1 RNase activity is tightly controlled by both intramolecular (NTD-PIN) and intermolecular (PIN-PIN) interactions. ABSTRACT | |
| 57 86 pattern-recognition receptors protein_type The initial sensing of infection is mediated by a set of pattern-recognition receptors (PRRs) such Toll-like receptors (TLRs) and the intracellular signaling cascades triggered by TLRs evoke transcriptional expression of inflammatory mediators that coordinate the elimination of pathogens and infected cells. INTRO | |
| 88 92 PRRs protein_type The initial sensing of infection is mediated by a set of pattern-recognition receptors (PRRs) such Toll-like receptors (TLRs) and the intracellular signaling cascades triggered by TLRs evoke transcriptional expression of inflammatory mediators that coordinate the elimination of pathogens and infected cells. INTRO | |
| 99 118 Toll-like receptors protein_type The initial sensing of infection is mediated by a set of pattern-recognition receptors (PRRs) such Toll-like receptors (TLRs) and the intracellular signaling cascades triggered by TLRs evoke transcriptional expression of inflammatory mediators that coordinate the elimination of pathogens and infected cells. INTRO | |
| 120 124 TLRs protein_type The initial sensing of infection is mediated by a set of pattern-recognition receptors (PRRs) such Toll-like receptors (TLRs) and the intracellular signaling cascades triggered by TLRs evoke transcriptional expression of inflammatory mediators that coordinate the elimination of pathogens and infected cells. INTRO | |
| 180 184 TLRs protein_type The initial sensing of infection is mediated by a set of pattern-recognition receptors (PRRs) such Toll-like receptors (TLRs) and the intracellular signaling cascades triggered by TLRs evoke transcriptional expression of inflammatory mediators that coordinate the elimination of pathogens and infected cells. INTRO | |
| 0 9 Regnase-1 protein Regnase-1 (also known as Zc3h12a and MCPIP1) is an RNase whose expression level is stimulated by lipopolysaccharides and prevents autoimmune diseases by directly controlling the stability of mRNAs of inflammatory genes such as interleukin (IL)-6, IL-1β, IL-2, and IL-12p40. INTRO | |
| 25 32 Zc3h12a protein Regnase-1 (also known as Zc3h12a and MCPIP1) is an RNase whose expression level is stimulated by lipopolysaccharides and prevents autoimmune diseases by directly controlling the stability of mRNAs of inflammatory genes such as interleukin (IL)-6, IL-1β, IL-2, and IL-12p40. INTRO | |
| 37 43 MCPIP1 protein Regnase-1 (also known as Zc3h12a and MCPIP1) is an RNase whose expression level is stimulated by lipopolysaccharides and prevents autoimmune diseases by directly controlling the stability of mRNAs of inflammatory genes such as interleukin (IL)-6, IL-1β, IL-2, and IL-12p40. INTRO | |
| 51 56 RNase protein_type Regnase-1 (also known as Zc3h12a and MCPIP1) is an RNase whose expression level is stimulated by lipopolysaccharides and prevents autoimmune diseases by directly controlling the stability of mRNAs of inflammatory genes such as interleukin (IL)-6, IL-1β, IL-2, and IL-12p40. INTRO | |
| 97 116 lipopolysaccharides chemical Regnase-1 (also known as Zc3h12a and MCPIP1) is an RNase whose expression level is stimulated by lipopolysaccharides and prevents autoimmune diseases by directly controlling the stability of mRNAs of inflammatory genes such as interleukin (IL)-6, IL-1β, IL-2, and IL-12p40. INTRO | |
| 191 196 mRNAs chemical Regnase-1 (also known as Zc3h12a and MCPIP1) is an RNase whose expression level is stimulated by lipopolysaccharides and prevents autoimmune diseases by directly controlling the stability of mRNAs of inflammatory genes such as interleukin (IL)-6, IL-1β, IL-2, and IL-12p40. INTRO | |
| 239 245 (IL)-6 protein_type Regnase-1 (also known as Zc3h12a and MCPIP1) is an RNase whose expression level is stimulated by lipopolysaccharides and prevents autoimmune diseases by directly controlling the stability of mRNAs of inflammatory genes such as interleukin (IL)-6, IL-1β, IL-2, and IL-12p40. INTRO | |
| 247 252 IL-1β protein_type Regnase-1 (also known as Zc3h12a and MCPIP1) is an RNase whose expression level is stimulated by lipopolysaccharides and prevents autoimmune diseases by directly controlling the stability of mRNAs of inflammatory genes such as interleukin (IL)-6, IL-1β, IL-2, and IL-12p40. INTRO | |
| 254 258 IL-2 protein_type Regnase-1 (also known as Zc3h12a and MCPIP1) is an RNase whose expression level is stimulated by lipopolysaccharides and prevents autoimmune diseases by directly controlling the stability of mRNAs of inflammatory genes such as interleukin (IL)-6, IL-1β, IL-2, and IL-12p40. INTRO | |
| 264 272 IL-12p40 protein_type Regnase-1 (also known as Zc3h12a and MCPIP1) is an RNase whose expression level is stimulated by lipopolysaccharides and prevents autoimmune diseases by directly controlling the stability of mRNAs of inflammatory genes such as interleukin (IL)-6, IL-1β, IL-2, and IL-12p40. INTRO | |
| 0 9 Regnase-1 protein Regnase-1 accelerates target mRNA degradation via their 3′-terminal untranslated region (3′UTR), and also degrades its own mRNA. INTRO | |
| 29 33 mRNA chemical Regnase-1 accelerates target mRNA degradation via their 3′-terminal untranslated region (3′UTR), and also degrades its own mRNA. INTRO | |
| 56 87 3′-terminal untranslated region structure_element Regnase-1 accelerates target mRNA degradation via their 3′-terminal untranslated region (3′UTR), and also degrades its own mRNA. INTRO | |
| 89 94 3′UTR structure_element Regnase-1 accelerates target mRNA degradation via their 3′-terminal untranslated region (3′UTR), and also degrades its own mRNA. INTRO | |
| 123 127 mRNA chemical Regnase-1 accelerates target mRNA degradation via their 3′-terminal untranslated region (3′UTR), and also degrades its own mRNA. INTRO | |
| 0 9 Regnase-1 protein Regnase-1 is a member of Regnase family and is composed of a PilT N-terminus like (PIN) domain followed by a CCCH-type zinc–finger (ZF) domain, which are conserved among Regnase family members. INTRO | |
| 25 39 Regnase family protein_type Regnase-1 is a member of Regnase family and is composed of a PilT N-terminus like (PIN) domain followed by a CCCH-type zinc–finger (ZF) domain, which are conserved among Regnase family members. INTRO | |
| 61 81 PilT N-terminus like structure_element Regnase-1 is a member of Regnase family and is composed of a PilT N-terminus like (PIN) domain followed by a CCCH-type zinc–finger (ZF) domain, which are conserved among Regnase family members. INTRO | |
| 83 86 PIN structure_element Regnase-1 is a member of Regnase family and is composed of a PilT N-terminus like (PIN) domain followed by a CCCH-type zinc–finger (ZF) domain, which are conserved among Regnase family members. INTRO | |
| 109 130 CCCH-type zinc–finger structure_element Regnase-1 is a member of Regnase family and is composed of a PilT N-terminus like (PIN) domain followed by a CCCH-type zinc–finger (ZF) domain, which are conserved among Regnase family members. INTRO | |
| 132 134 ZF structure_element Regnase-1 is a member of Regnase family and is composed of a PilT N-terminus like (PIN) domain followed by a CCCH-type zinc–finger (ZF) domain, which are conserved among Regnase family members. INTRO | |
| 154 163 conserved protein_state Regnase-1 is a member of Regnase family and is composed of a PilT N-terminus like (PIN) domain followed by a CCCH-type zinc–finger (ZF) domain, which are conserved among Regnase family members. INTRO | |
| 170 192 Regnase family members protein_type Regnase-1 is a member of Regnase family and is composed of a PilT N-terminus like (PIN) domain followed by a CCCH-type zinc–finger (ZF) domain, which are conserved among Regnase family members. INTRO | |
| 14 31 crystal structure evidence Recently, the crystal structure of the Regnase-1 PIN domain derived from Homo sapiens was reported. INTRO | |
| 39 48 Regnase-1 protein Recently, the crystal structure of the Regnase-1 PIN domain derived from Homo sapiens was reported. INTRO | |
| 49 52 PIN structure_element Recently, the crystal structure of the Regnase-1 PIN domain derived from Homo sapiens was reported. INTRO | |
| 73 85 Homo sapiens species Recently, the crystal structure of the Regnase-1 PIN domain derived from Homo sapiens was reported. INTRO | |
| 4 13 structure evidence The structure combined with functional analyses revealed that four catalytically important Asp residues form the catalytic center and stabilize Mg2+ binding that is crucial for RNase activity. INTRO | |
| 91 94 Asp residue_name The structure combined with functional analyses revealed that four catalytically important Asp residues form the catalytic center and stabilize Mg2+ binding that is crucial for RNase activity. INTRO | |
| 113 129 catalytic center site The structure combined with functional analyses revealed that four catalytically important Asp residues form the catalytic center and stabilize Mg2+ binding that is crucial for RNase activity. INTRO | |
| 144 148 Mg2+ chemical The structure combined with functional analyses revealed that four catalytically important Asp residues form the catalytic center and stabilize Mg2+ binding that is crucial for RNase activity. INTRO | |
| 177 182 RNase protein_type The structure combined with functional analyses revealed that four catalytically important Asp residues form the catalytic center and stabilize Mg2+ binding that is crucial for RNase activity. INTRO | |
| 8 27 CCCH-type ZF motifs structure_element Several CCCH-type ZF motifs in RNA-binding proteins have been reported to directly bind RNA. INTRO | |
| 31 51 RNA-binding proteins protein_type Several CCCH-type ZF motifs in RNA-binding proteins have been reported to directly bind RNA. INTRO | |
| 88 91 RNA chemical Several CCCH-type ZF motifs in RNA-binding proteins have been reported to directly bind RNA. INTRO | |
| 13 22 Regnase-1 protein In addition, Regnase-1 has been predicted to possess other domains in the N- and C- terminal regions. INTRO | |
| 74 100 N- and C- terminal regions structure_element In addition, Regnase-1 has been predicted to possess other domains in the N- and C- terminal regions. INTRO | |
| 13 22 structure evidence However, the structure and function of the ZF domain, N-terminal domain (NTD) and C-terminal domain (CTD) of Regnase-1 have not been solved. INTRO | |
| 43 45 ZF structure_element However, the structure and function of the ZF domain, N-terminal domain (NTD) and C-terminal domain (CTD) of Regnase-1 have not been solved. INTRO | |
| 54 71 N-terminal domain structure_element However, the structure and function of the ZF domain, N-terminal domain (NTD) and C-terminal domain (CTD) of Regnase-1 have not been solved. INTRO | |
| 73 76 NTD structure_element However, the structure and function of the ZF domain, N-terminal domain (NTD) and C-terminal domain (CTD) of Regnase-1 have not been solved. INTRO | |
| 82 99 C-terminal domain structure_element However, the structure and function of the ZF domain, N-terminal domain (NTD) and C-terminal domain (CTD) of Regnase-1 have not been solved. INTRO | |
| 101 104 CTD structure_element However, the structure and function of the ZF domain, N-terminal domain (NTD) and C-terminal domain (CTD) of Regnase-1 have not been solved. INTRO | |
| 109 118 Regnase-1 protein However, the structure and function of the ZF domain, N-terminal domain (NTD) and C-terminal domain (CTD) of Regnase-1 have not been solved. INTRO | |
| 19 53 structural and functional analyses experimental_method Here, we performed structural and functional analyses of individual domains of Regnase-1 derived from Mus musculus in order to understand the catalytic activity in vitro. INTRO | |
| 79 88 Regnase-1 protein Here, we performed structural and functional analyses of individual domains of Regnase-1 derived from Mus musculus in order to understand the catalytic activity in vitro. INTRO | |
| 102 114 Mus musculus species Here, we performed structural and functional analyses of individual domains of Regnase-1 derived from Mus musculus in order to understand the catalytic activity in vitro. INTRO | |
| 49 58 Regnase-1 protein Our data revealed that the catalytic activity of Regnase-1 is regulated through both intra and intermolecular domain interactions in vitro. INTRO | |
| 4 7 NTD structure_element The NTD plays a crucial role in efficient cleavage of target mRNA, through intramolecular NTD-PIN interactions. INTRO | |
| 61 65 mRNA chemical The NTD plays a crucial role in efficient cleavage of target mRNA, through intramolecular NTD-PIN interactions. INTRO | |
| 90 93 NTD structure_element The NTD plays a crucial role in efficient cleavage of target mRNA, through intramolecular NTD-PIN interactions. INTRO | |
| 94 97 PIN structure_element The NTD plays a crucial role in efficient cleavage of target mRNA, through intramolecular NTD-PIN interactions. INTRO | |
| 10 19 Regnase-1 protein Moreover, Regnase-1 functions as a dimer through intermolecular PIN-PIN interactions during cleavage of target mRNA. INTRO | |
| 35 40 dimer oligomeric_state Moreover, Regnase-1 functions as a dimer through intermolecular PIN-PIN interactions during cleavage of target mRNA. INTRO | |
| 64 67 PIN structure_element Moreover, Regnase-1 functions as a dimer through intermolecular PIN-PIN interactions during cleavage of target mRNA. INTRO | |
| 68 71 PIN structure_element Moreover, Regnase-1 functions as a dimer through intermolecular PIN-PIN interactions during cleavage of target mRNA. INTRO | |
| 111 115 mRNA chemical Moreover, Regnase-1 functions as a dimer through intermolecular PIN-PIN interactions during cleavage of target mRNA. INTRO | |
| 26 35 Regnase-1 protein Our findings suggest that Regnase-1 cleaves its target mRNA by an NTD-activated functional PIN dimer, while the ZF increases RNA affinity in the vicinity of the PIN dimer. INTRO | |
| 55 59 mRNA chemical Our findings suggest that Regnase-1 cleaves its target mRNA by an NTD-activated functional PIN dimer, while the ZF increases RNA affinity in the vicinity of the PIN dimer. INTRO | |
| 66 79 NTD-activated protein_state Our findings suggest that Regnase-1 cleaves its target mRNA by an NTD-activated functional PIN dimer, while the ZF increases RNA affinity in the vicinity of the PIN dimer. INTRO | |
| 80 90 functional protein_state Our findings suggest that Regnase-1 cleaves its target mRNA by an NTD-activated functional PIN dimer, while the ZF increases RNA affinity in the vicinity of the PIN dimer. INTRO | |
| 91 94 PIN structure_element Our findings suggest that Regnase-1 cleaves its target mRNA by an NTD-activated functional PIN dimer, while the ZF increases RNA affinity in the vicinity of the PIN dimer. INTRO | |
| 95 100 dimer oligomeric_state Our findings suggest that Regnase-1 cleaves its target mRNA by an NTD-activated functional PIN dimer, while the ZF increases RNA affinity in the vicinity of the PIN dimer. INTRO | |
| 112 114 ZF structure_element Our findings suggest that Regnase-1 cleaves its target mRNA by an NTD-activated functional PIN dimer, while the ZF increases RNA affinity in the vicinity of the PIN dimer. INTRO | |
| 125 128 RNA chemical Our findings suggest that Regnase-1 cleaves its target mRNA by an NTD-activated functional PIN dimer, while the ZF increases RNA affinity in the vicinity of the PIN dimer. INTRO | |
| 161 164 PIN structure_element Our findings suggest that Regnase-1 cleaves its target mRNA by an NTD-activated functional PIN dimer, while the ZF increases RNA affinity in the vicinity of the PIN dimer. INTRO | |
| 165 170 dimer oligomeric_state Our findings suggest that Regnase-1 cleaves its target mRNA by an NTD-activated functional PIN dimer, while the ZF increases RNA affinity in the vicinity of the PIN dimer. INTRO | |
| 7 17 structures evidence Domain structures of Regnase-1 RESULTS | |
| 21 30 Regnase-1 protein Domain structures of Regnase-1 RESULTS | |
| 12 21 Rengase-1 protein We analyzed Rengase-1 derived from Mus musculus and solved the structures of the four domains; NTD, PIN, ZF, and CTD individually by X-ray crystallography or NMR (Fig. 1a–e). RESULTS | |
| 35 47 Mus musculus species We analyzed Rengase-1 derived from Mus musculus and solved the structures of the four domains; NTD, PIN, ZF, and CTD individually by X-ray crystallography or NMR (Fig. 1a–e). RESULTS | |
| 52 58 solved experimental_method We analyzed Rengase-1 derived from Mus musculus and solved the structures of the four domains; NTD, PIN, ZF, and CTD individually by X-ray crystallography or NMR (Fig. 1a–e). RESULTS | |
| 63 73 structures evidence We analyzed Rengase-1 derived from Mus musculus and solved the structures of the four domains; NTD, PIN, ZF, and CTD individually by X-ray crystallography or NMR (Fig. 1a–e). RESULTS | |
| 95 98 NTD structure_element We analyzed Rengase-1 derived from Mus musculus and solved the structures of the four domains; NTD, PIN, ZF, and CTD individually by X-ray crystallography or NMR (Fig. 1a–e). RESULTS | |
| 100 103 PIN structure_element We analyzed Rengase-1 derived from Mus musculus and solved the structures of the four domains; NTD, PIN, ZF, and CTD individually by X-ray crystallography or NMR (Fig. 1a–e). RESULTS | |
| 105 107 ZF structure_element We analyzed Rengase-1 derived from Mus musculus and solved the structures of the four domains; NTD, PIN, ZF, and CTD individually by X-ray crystallography or NMR (Fig. 1a–e). RESULTS | |
| 113 116 CTD structure_element We analyzed Rengase-1 derived from Mus musculus and solved the structures of the four domains; NTD, PIN, ZF, and CTD individually by X-ray crystallography or NMR (Fig. 1a–e). RESULTS | |
| 133 154 X-ray crystallography experimental_method We analyzed Rengase-1 derived from Mus musculus and solved the structures of the four domains; NTD, PIN, ZF, and CTD individually by X-ray crystallography or NMR (Fig. 1a–e). RESULTS | |
| 158 161 NMR experimental_method We analyzed Rengase-1 derived from Mus musculus and solved the structures of the four domains; NTD, PIN, ZF, and CTD individually by X-ray crystallography or NMR (Fig. 1a–e). RESULTS | |
| 0 21 X-ray crystallography experimental_method X-ray crystallography was attempted for the fragment containing both the PIN and ZF domains, however, electron density was observed only for the PIN domain (Fig. 1c), consistent with a previous report on Regnase-1 derived from Homo sapiens. RESULTS | |
| 73 76 PIN structure_element X-ray crystallography was attempted for the fragment containing both the PIN and ZF domains, however, electron density was observed only for the PIN domain (Fig. 1c), consistent with a previous report on Regnase-1 derived from Homo sapiens. RESULTS | |
| 81 83 ZF structure_element X-ray crystallography was attempted for the fragment containing both the PIN and ZF domains, however, electron density was observed only for the PIN domain (Fig. 1c), consistent with a previous report on Regnase-1 derived from Homo sapiens. RESULTS | |
| 102 118 electron density evidence X-ray crystallography was attempted for the fragment containing both the PIN and ZF domains, however, electron density was observed only for the PIN domain (Fig. 1c), consistent with a previous report on Regnase-1 derived from Homo sapiens. RESULTS | |
| 145 148 PIN structure_element X-ray crystallography was attempted for the fragment containing both the PIN and ZF domains, however, electron density was observed only for the PIN domain (Fig. 1c), consistent with a previous report on Regnase-1 derived from Homo sapiens. RESULTS | |
| 204 213 Regnase-1 protein X-ray crystallography was attempted for the fragment containing both the PIN and ZF domains, however, electron density was observed only for the PIN domain (Fig. 1c), consistent with a previous report on Regnase-1 derived from Homo sapiens. RESULTS | |
| 227 239 Homo sapiens species X-ray crystallography was attempted for the fragment containing both the PIN and ZF domains, however, electron density was observed only for the PIN domain (Fig. 1c), consistent with a previous report on Regnase-1 derived from Homo sapiens. RESULTS | |
| 23 26 PIN structure_element This suggests that the PIN and ZF domains exist independently without interacting with each other. RESULTS | |
| 31 33 ZF structure_element This suggests that the PIN and ZF domains exist independently without interacting with each other. RESULTS | |
| 11 21 structures evidence The domain structures of NTD, ZF, and CTD were determined by NMR (Fig. 1b,d,e). RESULTS | |
| 25 28 NTD structure_element The domain structures of NTD, ZF, and CTD were determined by NMR (Fig. 1b,d,e). RESULTS | |
| 30 32 ZF structure_element The domain structures of NTD, ZF, and CTD were determined by NMR (Fig. 1b,d,e). RESULTS | |
| 38 41 CTD structure_element The domain structures of NTD, ZF, and CTD were determined by NMR (Fig. 1b,d,e). RESULTS | |
| 61 64 NMR experimental_method The domain structures of NTD, ZF, and CTD were determined by NMR (Fig. 1b,d,e). RESULTS | |
| 4 7 NTD structure_element The NTD and CTD are both composed of three α helices, and structurally resemble ubiquitin conjugating enzyme E2 K (PDB ID: 3K9O) and ubiquitin associated protein 1 (PDB ID: 4AE4), respectively, according to the Dali server. RESULTS | |
| 12 15 CTD structure_element The NTD and CTD are both composed of three α helices, and structurally resemble ubiquitin conjugating enzyme E2 K (PDB ID: 3K9O) and ubiquitin associated protein 1 (PDB ID: 4AE4), respectively, according to the Dali server. RESULTS | |
| 43 52 α helices structure_element The NTD and CTD are both composed of three α helices, and structurally resemble ubiquitin conjugating enzyme E2 K (PDB ID: 3K9O) and ubiquitin associated protein 1 (PDB ID: 4AE4), respectively, according to the Dali server. RESULTS | |
| 80 113 ubiquitin conjugating enzyme E2 K protein The NTD and CTD are both composed of three α helices, and structurally resemble ubiquitin conjugating enzyme E2 K (PDB ID: 3K9O) and ubiquitin associated protein 1 (PDB ID: 4AE4), respectively, according to the Dali server. RESULTS | |
| 133 163 ubiquitin associated protein 1 protein The NTD and CTD are both composed of three α helices, and structurally resemble ubiquitin conjugating enzyme E2 K (PDB ID: 3K9O) and ubiquitin associated protein 1 (PDB ID: 4AE4), respectively, according to the Dali server. RESULTS | |
| 211 222 Dali server experimental_method The NTD and CTD are both composed of three α helices, and structurally resemble ubiquitin conjugating enzyme E2 K (PDB ID: 3K9O) and ubiquitin associated protein 1 (PDB ID: 4AE4), respectively, according to the Dali server. RESULTS | |
| 31 40 Regnase-1 protein Contribution of each domain of Regnase-1 to the mRNA binding activity RESULTS | |
| 48 52 mRNA chemical Contribution of each domain of Regnase-1 to the mRNA binding activity RESULTS | |
| 13 16 PIN structure_element Although the PIN domain is responsible for the catalytic activity of Regnase-1, the roles of the other domains are largely unknown. RESULTS | |
| 69 78 Regnase-1 protein Although the PIN domain is responsible for the catalytic activity of Regnase-1, the roles of the other domains are largely unknown. RESULTS | |
| 34 37 NTD structure_element First, we evaluated a role of the NTD and ZF domains for mRNA binding by an in vitro gel shift assay (Fig. 1f). RESULTS | |
| 42 44 ZF structure_element First, we evaluated a role of the NTD and ZF domains for mRNA binding by an in vitro gel shift assay (Fig. 1f). RESULTS | |
| 57 61 mRNA chemical First, we evaluated a role of the NTD and ZF domains for mRNA binding by an in vitro gel shift assay (Fig. 1f). RESULTS | |
| 76 100 in vitro gel shift assay experimental_method First, we evaluated a role of the NTD and ZF domains for mRNA binding by an in vitro gel shift assay (Fig. 1f). RESULTS | |
| 0 24 Fluorescently 5′-labeled protein_state Fluorescently 5′-labeled RNA corresponding to nucleotides 82–106 of the IL-6 mRNA 3′UTR and the catalytically inactive mutant (D226N and D244N) of Regnase-1—hereafter referred to as the DDNN mutant—were utilized. RESULTS | |
| 25 28 RNA chemical Fluorescently 5′-labeled RNA corresponding to nucleotides 82–106 of the IL-6 mRNA 3′UTR and the catalytically inactive mutant (D226N and D244N) of Regnase-1—hereafter referred to as the DDNN mutant—were utilized. RESULTS | |
| 72 76 IL-6 protein_type Fluorescently 5′-labeled RNA corresponding to nucleotides 82–106 of the IL-6 mRNA 3′UTR and the catalytically inactive mutant (D226N and D244N) of Regnase-1—hereafter referred to as the DDNN mutant—were utilized. RESULTS | |
| 77 81 mRNA chemical Fluorescently 5′-labeled RNA corresponding to nucleotides 82–106 of the IL-6 mRNA 3′UTR and the catalytically inactive mutant (D226N and D244N) of Regnase-1—hereafter referred to as the DDNN mutant—were utilized. RESULTS | |
| 82 87 3′UTR structure_element Fluorescently 5′-labeled RNA corresponding to nucleotides 82–106 of the IL-6 mRNA 3′UTR and the catalytically inactive mutant (D226N and D244N) of Regnase-1—hereafter referred to as the DDNN mutant—were utilized. RESULTS | |
| 110 118 inactive protein_state Fluorescently 5′-labeled RNA corresponding to nucleotides 82–106 of the IL-6 mRNA 3′UTR and the catalytically inactive mutant (D226N and D244N) of Regnase-1—hereafter referred to as the DDNN mutant—were utilized. RESULTS | |
| 119 125 mutant protein_state Fluorescently 5′-labeled RNA corresponding to nucleotides 82–106 of the IL-6 mRNA 3′UTR and the catalytically inactive mutant (D226N and D244N) of Regnase-1—hereafter referred to as the DDNN mutant—were utilized. RESULTS | |
| 127 132 D226N mutant Fluorescently 5′-labeled RNA corresponding to nucleotides 82–106 of the IL-6 mRNA 3′UTR and the catalytically inactive mutant (D226N and D244N) of Regnase-1—hereafter referred to as the DDNN mutant—were utilized. RESULTS | |
| 137 142 D244N mutant Fluorescently 5′-labeled RNA corresponding to nucleotides 82–106 of the IL-6 mRNA 3′UTR and the catalytically inactive mutant (D226N and D244N) of Regnase-1—hereafter referred to as the DDNN mutant—were utilized. RESULTS | |
| 147 156 Regnase-1 protein Fluorescently 5′-labeled RNA corresponding to nucleotides 82–106 of the IL-6 mRNA 3′UTR and the catalytically inactive mutant (D226N and D244N) of Regnase-1—hereafter referred to as the DDNN mutant—were utilized. RESULTS | |
| 186 190 DDNN mutant Fluorescently 5′-labeled RNA corresponding to nucleotides 82–106 of the IL-6 mRNA 3′UTR and the catalytically inactive mutant (D226N and D244N) of Regnase-1—hereafter referred to as the DDNN mutant—were utilized. RESULTS | |
| 191 197 mutant protein_state Fluorescently 5′-labeled RNA corresponding to nucleotides 82–106 of the IL-6 mRNA 3′UTR and the catalytically inactive mutant (D226N and D244N) of Regnase-1—hereafter referred to as the DDNN mutant—were utilized. RESULTS | |
| 36 45 Regnase-1 protein Upon addition of a larger amount of Regnase-1, the fluorescence of free RNA decreased, indicating that Regnase-1 bound to the RNA. RESULTS | |
| 51 63 fluorescence evidence Upon addition of a larger amount of Regnase-1, the fluorescence of free RNA decreased, indicating that Regnase-1 bound to the RNA. RESULTS | |
| 67 71 free protein_state Upon addition of a larger amount of Regnase-1, the fluorescence of free RNA decreased, indicating that Regnase-1 bound to the RNA. RESULTS | |
| 72 75 RNA chemical Upon addition of a larger amount of Regnase-1, the fluorescence of free RNA decreased, indicating that Regnase-1 bound to the RNA. RESULTS | |
| 103 112 Regnase-1 protein Upon addition of a larger amount of Regnase-1, the fluorescence of free RNA decreased, indicating that Regnase-1 bound to the RNA. RESULTS | |
| 113 121 bound to protein_state Upon addition of a larger amount of Regnase-1, the fluorescence of free RNA decreased, indicating that Regnase-1 bound to the RNA. RESULTS | |
| 126 129 RNA chemical Upon addition of a larger amount of Regnase-1, the fluorescence of free RNA decreased, indicating that Regnase-1 bound to the RNA. RESULTS | |
| 34 37 RNA chemical Based on the decrease in the free RNA fluorescence band, we evaluated the contribution of each domain of Regnase-1 to RNA binding. RESULTS | |
| 105 114 Regnase-1 protein Based on the decrease in the free RNA fluorescence band, we evaluated the contribution of each domain of Regnase-1 to RNA binding. RESULTS | |
| 118 121 RNA chemical Based on the decrease in the free RNA fluorescence band, we evaluated the contribution of each domain of Regnase-1 to RNA binding. RESULTS | |
| 10 13 RNA chemical While the RNA binding ability was not significantly changed in the presence of NTD, it increased in the presence of the ZF domain (Fig. 1f,g and Supplementary Fig. 1). RESULTS | |
| 67 78 presence of protein_state While the RNA binding ability was not significantly changed in the presence of NTD, it increased in the presence of the ZF domain (Fig. 1f,g and Supplementary Fig. 1). RESULTS | |
| 79 82 NTD structure_element While the RNA binding ability was not significantly changed in the presence of NTD, it increased in the presence of the ZF domain (Fig. 1f,g and Supplementary Fig. 1). RESULTS | |
| 104 115 presence of protein_state While the RNA binding ability was not significantly changed in the presence of NTD, it increased in the presence of the ZF domain (Fig. 1f,g and Supplementary Fig. 1). RESULTS | |
| 120 122 ZF structure_element While the RNA binding ability was not significantly changed in the presence of NTD, it increased in the presence of the ZF domain (Fig. 1f,g and Supplementary Fig. 1). RESULTS | |
| 22 24 ZF structure_element Direct binding of the ZF domain and RNA were confirmed by NMR spectral changes. RESULTS | |
| 36 39 RNA chemical Direct binding of the ZF domain and RNA were confirmed by NMR spectral changes. RESULTS | |
| 58 61 NMR experimental_method Direct binding of the ZF domain and RNA were confirmed by NMR spectral changes. RESULTS | |
| 62 78 spectral changes evidence Direct binding of the ZF domain and RNA were confirmed by NMR spectral changes. RESULTS | |
| 19 34 titration curve evidence The fitting of the titration curve of Y314 resulted in an apparent dissociation constant (Kd) of 10 ± 1.1 μM (Supplementary Fig. 2). RESULTS | |
| 38 42 Y314 residue_name_number The fitting of the titration curve of Y314 resulted in an apparent dissociation constant (Kd) of 10 ± 1.1 μM (Supplementary Fig. 2). RESULTS | |
| 67 88 dissociation constant evidence The fitting of the titration curve of Y314 resulted in an apparent dissociation constant (Kd) of 10 ± 1.1 μM (Supplementary Fig. 2). RESULTS | |
| 90 92 Kd evidence The fitting of the titration curve of Y314 resulted in an apparent dissociation constant (Kd) of 10 ± 1.1 μM (Supplementary Fig. 2). RESULTS | |
| 41 44 PIN structure_element These results indicate that not only the PIN but also the ZF domain contribute to RNA binding, while the NTD is not likely to be involved in direct interaction with RNA. RESULTS | |
| 58 60 ZF structure_element These results indicate that not only the PIN but also the ZF domain contribute to RNA binding, while the NTD is not likely to be involved in direct interaction with RNA. RESULTS | |
| 82 85 RNA chemical These results indicate that not only the PIN but also the ZF domain contribute to RNA binding, while the NTD is not likely to be involved in direct interaction with RNA. RESULTS | |
| 105 108 NTD structure_element These results indicate that not only the PIN but also the ZF domain contribute to RNA binding, while the NTD is not likely to be involved in direct interaction with RNA. RESULTS | |
| 165 168 RNA chemical These results indicate that not only the PIN but also the ZF domain contribute to RNA binding, while the NTD is not likely to be involved in direct interaction with RNA. RESULTS | |
| 31 40 Regnase-1 protein Contribution of each domain of Regnase-1 to RNase activity RESULTS | |
| 44 49 RNase protein_type Contribution of each domain of Regnase-1 to RNase activity RESULTS | |
| 56 61 RNase protein_type In order to characterize the role of each domain in the RNase activity of Regnase-1, we performed an in vitro cleavage assay using fluorescently 5′-labeled RNA corresponding to nucleotides 82–106 of the IL-6 mRNA 3′UTR (Fig. 1g). RESULTS | |
| 74 83 Regnase-1 protein In order to characterize the role of each domain in the RNase activity of Regnase-1, we performed an in vitro cleavage assay using fluorescently 5′-labeled RNA corresponding to nucleotides 82–106 of the IL-6 mRNA 3′UTR (Fig. 1g). RESULTS | |
| 101 124 in vitro cleavage assay experimental_method In order to characterize the role of each domain in the RNase activity of Regnase-1, we performed an in vitro cleavage assay using fluorescently 5′-labeled RNA corresponding to nucleotides 82–106 of the IL-6 mRNA 3′UTR (Fig. 1g). RESULTS | |
| 131 155 fluorescently 5′-labeled protein_state In order to characterize the role of each domain in the RNase activity of Regnase-1, we performed an in vitro cleavage assay using fluorescently 5′-labeled RNA corresponding to nucleotides 82–106 of the IL-6 mRNA 3′UTR (Fig. 1g). RESULTS | |
| 156 159 RNA chemical In order to characterize the role of each domain in the RNase activity of Regnase-1, we performed an in vitro cleavage assay using fluorescently 5′-labeled RNA corresponding to nucleotides 82–106 of the IL-6 mRNA 3′UTR (Fig. 1g). RESULTS | |
| 203 207 IL-6 protein_type In order to characterize the role of each domain in the RNase activity of Regnase-1, we performed an in vitro cleavage assay using fluorescently 5′-labeled RNA corresponding to nucleotides 82–106 of the IL-6 mRNA 3′UTR (Fig. 1g). RESULTS | |
| 208 212 mRNA chemical In order to characterize the role of each domain in the RNase activity of Regnase-1, we performed an in vitro cleavage assay using fluorescently 5′-labeled RNA corresponding to nucleotides 82–106 of the IL-6 mRNA 3′UTR (Fig. 1g). RESULTS | |
| 213 218 3′UTR structure_element In order to characterize the role of each domain in the RNase activity of Regnase-1, we performed an in vitro cleavage assay using fluorescently 5′-labeled RNA corresponding to nucleotides 82–106 of the IL-6 mRNA 3′UTR (Fig. 1g). RESULTS | |
| 0 9 Regnase-1 protein Regnase-1 constructs consisting of NTD-PIN-ZF completely cleaved the target mRNA and generated the cleaved products. RESULTS | |
| 35 45 NTD-PIN-ZF mutant Regnase-1 constructs consisting of NTD-PIN-ZF completely cleaved the target mRNA and generated the cleaved products. RESULTS | |
| 76 80 mRNA chemical Regnase-1 constructs consisting of NTD-PIN-ZF completely cleaved the target mRNA and generated the cleaved products. RESULTS | |
| 37 42 RNase protein_type The apparent half-life (T1/2) of the RNase activity was about 20 minutes. RESULTS | |
| 0 9 Regnase-1 protein Regnase-1 lacking the ZF domain generated a smaller but appreciable amount of cleaved product (T1/2 ~ 70 minutes), while those lacking the NTD did not generate cleaved products (T1/2 > 90 minutes). RESULTS | |
| 10 17 lacking protein_state Regnase-1 lacking the ZF domain generated a smaller but appreciable amount of cleaved product (T1/2 ~ 70 minutes), while those lacking the NTD did not generate cleaved products (T1/2 > 90 minutes). RESULTS | |
| 22 24 ZF structure_element Regnase-1 lacking the ZF domain generated a smaller but appreciable amount of cleaved product (T1/2 ~ 70 minutes), while those lacking the NTD did not generate cleaved products (T1/2 > 90 minutes). RESULTS | |
| 127 134 lacking protein_state Regnase-1 lacking the ZF domain generated a smaller but appreciable amount of cleaved product (T1/2 ~ 70 minutes), while those lacking the NTD did not generate cleaved products (T1/2 > 90 minutes). RESULTS | |
| 139 142 NTD structure_element Regnase-1 lacking the ZF domain generated a smaller but appreciable amount of cleaved product (T1/2 ~ 70 minutes), while those lacking the NTD did not generate cleaved products (T1/2 > 90 minutes). RESULTS | |
| 24 40 NTD-PIN(DDNN)-ZF mutant It should be noted that NTD-PIN(DDNN)-ZF, which possesses the NTD but lacks the catalytic residues in PIN, completely lost all RNase activity (Fig. 1g, right panel), as expected, confirming that the RNase catalytic center is located in the PIN domain. RESULTS | |
| 62 65 NTD structure_element It should be noted that NTD-PIN(DDNN)-ZF, which possesses the NTD but lacks the catalytic residues in PIN, completely lost all RNase activity (Fig. 1g, right panel), as expected, confirming that the RNase catalytic center is located in the PIN domain. RESULTS | |
| 70 75 lacks protein_state It should be noted that NTD-PIN(DDNN)-ZF, which possesses the NTD but lacks the catalytic residues in PIN, completely lost all RNase activity (Fig. 1g, right panel), as expected, confirming that the RNase catalytic center is located in the PIN domain. RESULTS | |
| 80 98 catalytic residues site It should be noted that NTD-PIN(DDNN)-ZF, which possesses the NTD but lacks the catalytic residues in PIN, completely lost all RNase activity (Fig. 1g, right panel), as expected, confirming that the RNase catalytic center is located in the PIN domain. RESULTS | |
| 102 105 PIN structure_element It should be noted that NTD-PIN(DDNN)-ZF, which possesses the NTD but lacks the catalytic residues in PIN, completely lost all RNase activity (Fig. 1g, right panel), as expected, confirming that the RNase catalytic center is located in the PIN domain. RESULTS | |
| 127 132 RNase protein_type It should be noted that NTD-PIN(DDNN)-ZF, which possesses the NTD but lacks the catalytic residues in PIN, completely lost all RNase activity (Fig. 1g, right panel), as expected, confirming that the RNase catalytic center is located in the PIN domain. RESULTS | |
| 199 204 RNase protein_type It should be noted that NTD-PIN(DDNN)-ZF, which possesses the NTD but lacks the catalytic residues in PIN, completely lost all RNase activity (Fig. 1g, right panel), as expected, confirming that the RNase catalytic center is located in the PIN domain. RESULTS | |
| 205 221 catalytic center site It should be noted that NTD-PIN(DDNN)-ZF, which possesses the NTD but lacks the catalytic residues in PIN, completely lost all RNase activity (Fig. 1g, right panel), as expected, confirming that the RNase catalytic center is located in the PIN domain. RESULTS | |
| 240 243 PIN structure_element It should be noted that NTD-PIN(DDNN)-ZF, which possesses the NTD but lacks the catalytic residues in PIN, completely lost all RNase activity (Fig. 1g, right panel), as expected, confirming that the RNase catalytic center is located in the PIN domain. RESULTS | |
| 78 81 NTD structure_element Taken together with the results in the previous section, we conclude that the NTD is crucial for the RNase activity of Regnase-1 in vitro, although it does not contribute to the direct mRNA binding. RESULTS | |
| 101 106 RNase protein_type Taken together with the results in the previous section, we conclude that the NTD is crucial for the RNase activity of Regnase-1 in vitro, although it does not contribute to the direct mRNA binding. RESULTS | |
| 119 128 Regnase-1 protein Taken together with the results in the previous section, we conclude that the NTD is crucial for the RNase activity of Regnase-1 in vitro, although it does not contribute to the direct mRNA binding. RESULTS | |
| 185 189 mRNA chemical Taken together with the results in the previous section, we conclude that the NTD is crucial for the RNase activity of Regnase-1 in vitro, although it does not contribute to the direct mRNA binding. RESULTS | |
| 0 5 Dimer oligomeric_state Dimer formation of the PIN domains RESULTS | |
| 23 26 PIN structure_element Dimer formation of the PIN domains RESULTS | |
| 7 19 purification experimental_method During purification by gel filtration, the PIN domain exhibited extremely asymmetric elution peaks in a concentration dependent manner (Fig. 2a). RESULTS | |
| 23 37 gel filtration experimental_method During purification by gel filtration, the PIN domain exhibited extremely asymmetric elution peaks in a concentration dependent manner (Fig. 2a). RESULTS | |
| 43 46 PIN structure_element During purification by gel filtration, the PIN domain exhibited extremely asymmetric elution peaks in a concentration dependent manner (Fig. 2a). RESULTS | |
| 3 65 comparison with the elution volume of standard marker proteins experimental_method By comparison with the elution volume of standard marker proteins, the PIN domain was assumed to be in equilibrium between a monomer and a dimer in solution at concentrations in the 20–200 μM range. RESULTS | |
| 71 74 PIN structure_element By comparison with the elution volume of standard marker proteins, the PIN domain was assumed to be in equilibrium between a monomer and a dimer in solution at concentrations in the 20–200 μM range. RESULTS | |
| 125 132 monomer oligomeric_state By comparison with the elution volume of standard marker proteins, the PIN domain was assumed to be in equilibrium between a monomer and a dimer in solution at concentrations in the 20–200 μM range. RESULTS | |
| 139 144 dimer oligomeric_state By comparison with the elution volume of standard marker proteins, the PIN domain was assumed to be in equilibrium between a monomer and a dimer in solution at concentrations in the 20–200 μM range. RESULTS | |
| 4 21 crystal structure evidence The crystal structure of the PIN domain has been determined in three distinct crystal forms with a space group of P3121 (form I in this study and PDB ID 3V33), P3221 (form II in this study), and P41 (PDB ID 3V32 and 3V34), respectively. RESULTS | |
| 29 32 PIN structure_element The crystal structure of the PIN domain has been determined in three distinct crystal forms with a space group of P3121 (form I in this study and PDB ID 3V33), P3221 (form II in this study), and P41 (PDB ID 3V32 and 3V34), respectively. RESULTS | |
| 78 91 crystal forms evidence The crystal structure of the PIN domain has been determined in three distinct crystal forms with a space group of P3121 (form I in this study and PDB ID 3V33), P3221 (form II in this study), and P41 (PDB ID 3V32 and 3V34), respectively. RESULTS | |
| 18 21 PIN structure_element We found that the PIN domain formed a head-to-tail oligomer that was commonly observed in all three crystal forms in spite of the different crystallization conditions (Supplementary Fig. 3). RESULTS | |
| 38 50 head-to-tail protein_state We found that the PIN domain formed a head-to-tail oligomer that was commonly observed in all three crystal forms in spite of the different crystallization conditions (Supplementary Fig. 3). RESULTS | |
| 51 59 oligomer oligomeric_state We found that the PIN domain formed a head-to-tail oligomer that was commonly observed in all three crystal forms in spite of the different crystallization conditions (Supplementary Fig. 3). RESULTS | |
| 100 113 crystal forms evidence We found that the PIN domain formed a head-to-tail oligomer that was commonly observed in all three crystal forms in spite of the different crystallization conditions (Supplementary Fig. 3). RESULTS | |
| 0 8 Mutation experimental_method Mutation of Arg215, whose side chain faces to the opposite side of the oligomeric surface, to Glu preserved the monomer/dimer equilibrium, similar to the wild type. RESULTS | |
| 12 18 Arg215 residue_name_number Mutation of Arg215, whose side chain faces to the opposite side of the oligomeric surface, to Glu preserved the monomer/dimer equilibrium, similar to the wild type. RESULTS | |
| 71 89 oligomeric surface site Mutation of Arg215, whose side chain faces to the opposite side of the oligomeric surface, to Glu preserved the monomer/dimer equilibrium, similar to the wild type. RESULTS | |
| 94 97 Glu residue_name Mutation of Arg215, whose side chain faces to the opposite side of the oligomeric surface, to Glu preserved the monomer/dimer equilibrium, similar to the wild type. RESULTS | |
| 112 119 monomer oligomeric_state Mutation of Arg215, whose side chain faces to the opposite side of the oligomeric surface, to Glu preserved the monomer/dimer equilibrium, similar to the wild type. RESULTS | |
| 120 125 dimer oligomeric_state Mutation of Arg215, whose side chain faces to the opposite side of the oligomeric surface, to Glu preserved the monomer/dimer equilibrium, similar to the wild type. RESULTS | |
| 154 163 wild type protein_state Mutation of Arg215, whose side chain faces to the opposite side of the oligomeric surface, to Glu preserved the monomer/dimer equilibrium, similar to the wild type. RESULTS | |
| 19 35 single mutations experimental_method On the other hand, single mutations of side chains involved in the PIN–PIN oligomeric interaction resulted in monomer formation, judging from gel filtration (Fig. 2a,b). RESULTS | |
| 67 70 PIN structure_element On the other hand, single mutations of side chains involved in the PIN–PIN oligomeric interaction resulted in monomer formation, judging from gel filtration (Fig. 2a,b). RESULTS | |
| 71 74 PIN structure_element On the other hand, single mutations of side chains involved in the PIN–PIN oligomeric interaction resulted in monomer formation, judging from gel filtration (Fig. 2a,b). RESULTS | |
| 110 117 monomer oligomeric_state On the other hand, single mutations of side chains involved in the PIN–PIN oligomeric interaction resulted in monomer formation, judging from gel filtration (Fig. 2a,b). RESULTS | |
| 142 156 gel filtration experimental_method On the other hand, single mutations of side chains involved in the PIN–PIN oligomeric interaction resulted in monomer formation, judging from gel filtration (Fig. 2a,b). RESULTS | |
| 0 9 Wild type protein_state Wild type and monomeric PIN mutants (P212A and D278R) were also analyzed by NMR. RESULTS | |
| 14 23 monomeric oligomeric_state Wild type and monomeric PIN mutants (P212A and D278R) were also analyzed by NMR. RESULTS | |
| 24 27 PIN structure_element Wild type and monomeric PIN mutants (P212A and D278R) were also analyzed by NMR. RESULTS | |
| 28 35 mutants protein_state Wild type and monomeric PIN mutants (P212A and D278R) were also analyzed by NMR. RESULTS | |
| 37 42 P212A mutant Wild type and monomeric PIN mutants (P212A and D278R) were also analyzed by NMR. RESULTS | |
| 47 52 D278R mutant Wild type and monomeric PIN mutants (P212A and D278R) were also analyzed by NMR. RESULTS | |
| 76 79 NMR experimental_method Wild type and monomeric PIN mutants (P212A and D278R) were also analyzed by NMR. RESULTS | |
| 4 11 spectra evidence The spectra indicate that the dimer interface of the wild type PIN domain were significantly broadened compared to the monomeric mutants (Supplementary Fig. 4). RESULTS | |
| 30 45 dimer interface site The spectra indicate that the dimer interface of the wild type PIN domain were significantly broadened compared to the monomeric mutants (Supplementary Fig. 4). RESULTS | |
| 53 62 wild type protein_state The spectra indicate that the dimer interface of the wild type PIN domain were significantly broadened compared to the monomeric mutants (Supplementary Fig. 4). RESULTS | |
| 63 66 PIN structure_element The spectra indicate that the dimer interface of the wild type PIN domain were significantly broadened compared to the monomeric mutants (Supplementary Fig. 4). RESULTS | |
| 119 128 monomeric oligomeric_state The spectra indicate that the dimer interface of the wild type PIN domain were significantly broadened compared to the monomeric mutants (Supplementary Fig. 4). RESULTS | |
| 129 136 mutants protein_state The spectra indicate that the dimer interface of the wild type PIN domain were significantly broadened compared to the monomeric mutants (Supplementary Fig. 4). RESULTS | |
| 32 35 PIN structure_element These results indicate that the PIN domain forms a head-to-tail oligomer in solution similar to the crystal structure. RESULTS | |
| 51 63 head-to-tail protein_state These results indicate that the PIN domain forms a head-to-tail oligomer in solution similar to the crystal structure. RESULTS | |
| 64 72 oligomer oligomeric_state These results indicate that the PIN domain forms a head-to-tail oligomer in solution similar to the crystal structure. RESULTS | |
| 100 117 crystal structure evidence These results indicate that the PIN domain forms a head-to-tail oligomer in solution similar to the crystal structure. RESULTS | |
| 19 28 monomeric oligomeric_state Interestingly, the monomeric PIN mutants P212A, R214A, and D278R had no significant RNase activity for IL-6 mRNA in vitro (Fig. 2c). RESULTS | |
| 29 32 PIN structure_element Interestingly, the monomeric PIN mutants P212A, R214A, and D278R had no significant RNase activity for IL-6 mRNA in vitro (Fig. 2c). RESULTS | |
| 33 40 mutants protein_state Interestingly, the monomeric PIN mutants P212A, R214A, and D278R had no significant RNase activity for IL-6 mRNA in vitro (Fig. 2c). RESULTS | |
| 41 46 P212A mutant Interestingly, the monomeric PIN mutants P212A, R214A, and D278R had no significant RNase activity for IL-6 mRNA in vitro (Fig. 2c). RESULTS | |
| 48 53 R214A mutant Interestingly, the monomeric PIN mutants P212A, R214A, and D278R had no significant RNase activity for IL-6 mRNA in vitro (Fig. 2c). RESULTS | |
| 59 64 D278R mutant Interestingly, the monomeric PIN mutants P212A, R214A, and D278R had no significant RNase activity for IL-6 mRNA in vitro (Fig. 2c). RESULTS | |
| 84 89 RNase protein_type Interestingly, the monomeric PIN mutants P212A, R214A, and D278R had no significant RNase activity for IL-6 mRNA in vitro (Fig. 2c). RESULTS | |
| 103 107 IL-6 protein_type Interestingly, the monomeric PIN mutants P212A, R214A, and D278R had no significant RNase activity for IL-6 mRNA in vitro (Fig. 2c). RESULTS | |
| 108 112 mRNA chemical Interestingly, the monomeric PIN mutants P212A, R214A, and D278R had no significant RNase activity for IL-6 mRNA in vitro (Fig. 2c). RESULTS | |
| 54 70 catalytic center site The side chains of these residues point away from the catalytic center on the same molecule (Fig. 2b). RESULTS | |
| 29 41 head-to-tail protein_state Therefore, we concluded that head-to-tail PIN dimerization, together with the NTD, are required for Regnase-1 RNase activity in vitro. RESULTS | |
| 42 45 PIN structure_element Therefore, we concluded that head-to-tail PIN dimerization, together with the NTD, are required for Regnase-1 RNase activity in vitro. RESULTS | |
| 78 81 NTD structure_element Therefore, we concluded that head-to-tail PIN dimerization, together with the NTD, are required for Regnase-1 RNase activity in vitro. RESULTS | |
| 100 109 Regnase-1 protein Therefore, we concluded that head-to-tail PIN dimerization, together with the NTD, are required for Regnase-1 RNase activity in vitro. RESULTS | |
| 110 115 RNase protein_type Therefore, we concluded that head-to-tail PIN dimerization, together with the NTD, are required for Regnase-1 RNase activity in vitro. RESULTS | |
| 38 41 NTD structure_element Domain-domain interaction between the NTD and the PIN domain RESULTS | |
| 50 53 PIN structure_element Domain-domain interaction between the NTD and the PIN domain RESULTS | |
| 10 13 NTD structure_element While the NTD does not contribute to RNA binding (Fig. 1f,g, and Supplementary Fig. 1), it increases the RNase activity of Regnase-1 (Fig. 1h). RESULTS | |
| 37 40 RNA chemical While the NTD does not contribute to RNA binding (Fig. 1f,g, and Supplementary Fig. 1), it increases the RNase activity of Regnase-1 (Fig. 1h). RESULTS | |
| 105 110 RNase protein_type While the NTD does not contribute to RNA binding (Fig. 1f,g, and Supplementary Fig. 1), it increases the RNase activity of Regnase-1 (Fig. 1h). RESULTS | |
| 123 132 Regnase-1 protein While the NTD does not contribute to RNA binding (Fig. 1f,g, and Supplementary Fig. 1), it increases the RNase activity of Regnase-1 (Fig. 1h). RESULTS | |
| 61 64 NTD structure_element In order to gain insight into the molecular mechanism of the NTD-mediated enhancement of Regnase-1 RNase activity, we further investigated the domain-domain interaction between the NTD and the PIN domain using NMR. RESULTS | |
| 89 98 Regnase-1 protein In order to gain insight into the molecular mechanism of the NTD-mediated enhancement of Regnase-1 RNase activity, we further investigated the domain-domain interaction between the NTD and the PIN domain using NMR. RESULTS | |
| 99 104 RNase protein_type In order to gain insight into the molecular mechanism of the NTD-mediated enhancement of Regnase-1 RNase activity, we further investigated the domain-domain interaction between the NTD and the PIN domain using NMR. RESULTS | |
| 181 184 NTD structure_element In order to gain insight into the molecular mechanism of the NTD-mediated enhancement of Regnase-1 RNase activity, we further investigated the domain-domain interaction between the NTD and the PIN domain using NMR. RESULTS | |
| 193 196 PIN structure_element In order to gain insight into the molecular mechanism of the NTD-mediated enhancement of Regnase-1 RNase activity, we further investigated the domain-domain interaction between the NTD and the PIN domain using NMR. RESULTS | |
| 210 213 NMR experimental_method In order to gain insight into the molecular mechanism of the NTD-mediated enhancement of Regnase-1 RNase activity, we further investigated the domain-domain interaction between the NTD and the PIN domain using NMR. RESULTS | |
| 12 34 catalytically inactive protein_state We used the catalytically inactive monomeric PIN mutant possessing both the DDNN and D278R mutations to avoid dimer formation of the PIN domain. RESULTS | |
| 35 44 monomeric oligomeric_state We used the catalytically inactive monomeric PIN mutant possessing both the DDNN and D278R mutations to avoid dimer formation of the PIN domain. RESULTS | |
| 45 48 PIN structure_element We used the catalytically inactive monomeric PIN mutant possessing both the DDNN and D278R mutations to avoid dimer formation of the PIN domain. RESULTS | |
| 49 55 mutant protein_state We used the catalytically inactive monomeric PIN mutant possessing both the DDNN and D278R mutations to avoid dimer formation of the PIN domain. RESULTS | |
| 76 80 DDNN mutant We used the catalytically inactive monomeric PIN mutant possessing both the DDNN and D278R mutations to avoid dimer formation of the PIN domain. RESULTS | |
| 85 90 D278R mutant We used the catalytically inactive monomeric PIN mutant possessing both the DDNN and D278R mutations to avoid dimer formation of the PIN domain. RESULTS | |
| 110 115 dimer oligomeric_state We used the catalytically inactive monomeric PIN mutant possessing both the DDNN and D278R mutations to avoid dimer formation of the PIN domain. RESULTS | |
| 133 136 PIN structure_element We used the catalytically inactive monomeric PIN mutant possessing both the DDNN and D278R mutations to avoid dimer formation of the PIN domain. RESULTS | |
| 4 7 NMR experimental_method The NMR signals from the PIN domain (residues V177, F210-T211, R214, F228-L232, and F234-S236) exhibited significant chemical shift changes upon addition of the NTD (Fig. 3a). RESULTS | |
| 25 28 PIN structure_element The NMR signals from the PIN domain (residues V177, F210-T211, R214, F228-L232, and F234-S236) exhibited significant chemical shift changes upon addition of the NTD (Fig. 3a). RESULTS | |
| 46 50 V177 residue_name_number The NMR signals from the PIN domain (residues V177, F210-T211, R214, F228-L232, and F234-S236) exhibited significant chemical shift changes upon addition of the NTD (Fig. 3a). RESULTS | |
| 52 61 F210-T211 residue_range The NMR signals from the PIN domain (residues V177, F210-T211, R214, F228-L232, and F234-S236) exhibited significant chemical shift changes upon addition of the NTD (Fig. 3a). RESULTS | |
| 63 67 R214 residue_name_number The NMR signals from the PIN domain (residues V177, F210-T211, R214, F228-L232, and F234-S236) exhibited significant chemical shift changes upon addition of the NTD (Fig. 3a). RESULTS | |
| 69 78 F228-L232 residue_range The NMR signals from the PIN domain (residues V177, F210-T211, R214, F228-L232, and F234-S236) exhibited significant chemical shift changes upon addition of the NTD (Fig. 3a). RESULTS | |
| 84 93 F234-S236 residue_range The NMR signals from the PIN domain (residues V177, F210-T211, R214, F228-L232, and F234-S236) exhibited significant chemical shift changes upon addition of the NTD (Fig. 3a). RESULTS | |
| 145 156 addition of experimental_method The NMR signals from the PIN domain (residues V177, F210-T211, R214, F228-L232, and F234-S236) exhibited significant chemical shift changes upon addition of the NTD (Fig. 3a). RESULTS | |
| 161 164 NTD structure_element The NMR signals from the PIN domain (residues V177, F210-T211, R214, F228-L232, and F234-S236) exhibited significant chemical shift changes upon addition of the NTD (Fig. 3a). RESULTS | |
| 15 26 addition of experimental_method Likewise, upon addition of the PIN domain, NMR signals derived from R56, L58-G59, and V86-H88 in the NTD exhibited large chemical shift changes and residues D53, F55, K57, Y60-S61, V68, T80-G83, L85, and G89 of the NTD as well as side chain amide signals of N79 exhibited small but appreciable chemical shift changes (Fig. 3b and Supplementary Fig. 5). RESULTS | |
| 31 34 PIN structure_element Likewise, upon addition of the PIN domain, NMR signals derived from R56, L58-G59, and V86-H88 in the NTD exhibited large chemical shift changes and residues D53, F55, K57, Y60-S61, V68, T80-G83, L85, and G89 of the NTD as well as side chain amide signals of N79 exhibited small but appreciable chemical shift changes (Fig. 3b and Supplementary Fig. 5). RESULTS | |
| 43 46 NMR experimental_method Likewise, upon addition of the PIN domain, NMR signals derived from R56, L58-G59, and V86-H88 in the NTD exhibited large chemical shift changes and residues D53, F55, K57, Y60-S61, V68, T80-G83, L85, and G89 of the NTD as well as side chain amide signals of N79 exhibited small but appreciable chemical shift changes (Fig. 3b and Supplementary Fig. 5). RESULTS | |
| 68 71 R56 residue_name_number Likewise, upon addition of the PIN domain, NMR signals derived from R56, L58-G59, and V86-H88 in the NTD exhibited large chemical shift changes and residues D53, F55, K57, Y60-S61, V68, T80-G83, L85, and G89 of the NTD as well as side chain amide signals of N79 exhibited small but appreciable chemical shift changes (Fig. 3b and Supplementary Fig. 5). RESULTS | |
| 73 80 L58-G59 residue_range Likewise, upon addition of the PIN domain, NMR signals derived from R56, L58-G59, and V86-H88 in the NTD exhibited large chemical shift changes and residues D53, F55, K57, Y60-S61, V68, T80-G83, L85, and G89 of the NTD as well as side chain amide signals of N79 exhibited small but appreciable chemical shift changes (Fig. 3b and Supplementary Fig. 5). RESULTS | |
| 86 93 V86-H88 residue_range Likewise, upon addition of the PIN domain, NMR signals derived from R56, L58-G59, and V86-H88 in the NTD exhibited large chemical shift changes and residues D53, F55, K57, Y60-S61, V68, T80-G83, L85, and G89 of the NTD as well as side chain amide signals of N79 exhibited small but appreciable chemical shift changes (Fig. 3b and Supplementary Fig. 5). RESULTS | |
| 101 104 NTD structure_element Likewise, upon addition of the PIN domain, NMR signals derived from R56, L58-G59, and V86-H88 in the NTD exhibited large chemical shift changes and residues D53, F55, K57, Y60-S61, V68, T80-G83, L85, and G89 of the NTD as well as side chain amide signals of N79 exhibited small but appreciable chemical shift changes (Fig. 3b and Supplementary Fig. 5). RESULTS | |
| 157 160 D53 residue_name_number Likewise, upon addition of the PIN domain, NMR signals derived from R56, L58-G59, and V86-H88 in the NTD exhibited large chemical shift changes and residues D53, F55, K57, Y60-S61, V68, T80-G83, L85, and G89 of the NTD as well as side chain amide signals of N79 exhibited small but appreciable chemical shift changes (Fig. 3b and Supplementary Fig. 5). RESULTS | |
| 162 165 F55 residue_name_number Likewise, upon addition of the PIN domain, NMR signals derived from R56, L58-G59, and V86-H88 in the NTD exhibited large chemical shift changes and residues D53, F55, K57, Y60-S61, V68, T80-G83, L85, and G89 of the NTD as well as side chain amide signals of N79 exhibited small but appreciable chemical shift changes (Fig. 3b and Supplementary Fig. 5). RESULTS | |
| 167 170 K57 residue_name_number Likewise, upon addition of the PIN domain, NMR signals derived from R56, L58-G59, and V86-H88 in the NTD exhibited large chemical shift changes and residues D53, F55, K57, Y60-S61, V68, T80-G83, L85, and G89 of the NTD as well as side chain amide signals of N79 exhibited small but appreciable chemical shift changes (Fig. 3b and Supplementary Fig. 5). RESULTS | |
| 172 179 Y60-S61 residue_range Likewise, upon addition of the PIN domain, NMR signals derived from R56, L58-G59, and V86-H88 in the NTD exhibited large chemical shift changes and residues D53, F55, K57, Y60-S61, V68, T80-G83, L85, and G89 of the NTD as well as side chain amide signals of N79 exhibited small but appreciable chemical shift changes (Fig. 3b and Supplementary Fig. 5). RESULTS | |
| 181 184 V68 residue_name_number Likewise, upon addition of the PIN domain, NMR signals derived from R56, L58-G59, and V86-H88 in the NTD exhibited large chemical shift changes and residues D53, F55, K57, Y60-S61, V68, T80-G83, L85, and G89 of the NTD as well as side chain amide signals of N79 exhibited small but appreciable chemical shift changes (Fig. 3b and Supplementary Fig. 5). RESULTS | |
| 186 193 T80-G83 residue_range Likewise, upon addition of the PIN domain, NMR signals derived from R56, L58-G59, and V86-H88 in the NTD exhibited large chemical shift changes and residues D53, F55, K57, Y60-S61, V68, T80-G83, L85, and G89 of the NTD as well as side chain amide signals of N79 exhibited small but appreciable chemical shift changes (Fig. 3b and Supplementary Fig. 5). RESULTS | |
| 195 198 L85 residue_name_number Likewise, upon addition of the PIN domain, NMR signals derived from R56, L58-G59, and V86-H88 in the NTD exhibited large chemical shift changes and residues D53, F55, K57, Y60-S61, V68, T80-G83, L85, and G89 of the NTD as well as side chain amide signals of N79 exhibited small but appreciable chemical shift changes (Fig. 3b and Supplementary Fig. 5). RESULTS | |
| 204 207 G89 residue_name_number Likewise, upon addition of the PIN domain, NMR signals derived from R56, L58-G59, and V86-H88 in the NTD exhibited large chemical shift changes and residues D53, F55, K57, Y60-S61, V68, T80-G83, L85, and G89 of the NTD as well as side chain amide signals of N79 exhibited small but appreciable chemical shift changes (Fig. 3b and Supplementary Fig. 5). RESULTS | |
| 215 218 NTD structure_element Likewise, upon addition of the PIN domain, NMR signals derived from R56, L58-G59, and V86-H88 in the NTD exhibited large chemical shift changes and residues D53, F55, K57, Y60-S61, V68, T80-G83, L85, and G89 of the NTD as well as side chain amide signals of N79 exhibited small but appreciable chemical shift changes (Fig. 3b and Supplementary Fig. 5). RESULTS | |
| 258 261 N79 residue_name_number Likewise, upon addition of the PIN domain, NMR signals derived from R56, L58-G59, and V86-H88 in the NTD exhibited large chemical shift changes and residues D53, F55, K57, Y60-S61, V68, T80-G83, L85, and G89 of the NTD as well as side chain amide signals of N79 exhibited small but appreciable chemical shift changes (Fig. 3b and Supplementary Fig. 5). RESULTS | |
| 64 67 PIN structure_element These results clearly indicate a direct interaction between the PIN domain and the NTD. RESULTS | |
| 83 86 NTD structure_element These results clearly indicate a direct interaction between the PIN domain and the NTD. RESULTS | |
| 13 28 titration curve evidence Based on the titration curve for the chemical shift changes of L58, the apparent Kd between the isolated NTD and PIN was estimated to be 110 ± 5.8 μM. Considering the fact that the NTD and PIN domains are attached by a linker, the actual binding affinity is expected much higher in the native protein. RESULTS | |
| 37 59 chemical shift changes evidence Based on the titration curve for the chemical shift changes of L58, the apparent Kd between the isolated NTD and PIN was estimated to be 110 ± 5.8 μM. Considering the fact that the NTD and PIN domains are attached by a linker, the actual binding affinity is expected much higher in the native protein. RESULTS | |
| 63 66 L58 residue_name_number Based on the titration curve for the chemical shift changes of L58, the apparent Kd between the isolated NTD and PIN was estimated to be 110 ± 5.8 μM. Considering the fact that the NTD and PIN domains are attached by a linker, the actual binding affinity is expected much higher in the native protein. RESULTS | |
| 81 83 Kd evidence Based on the titration curve for the chemical shift changes of L58, the apparent Kd between the isolated NTD and PIN was estimated to be 110 ± 5.8 μM. Considering the fact that the NTD and PIN domains are attached by a linker, the actual binding affinity is expected much higher in the native protein. RESULTS | |
| 105 108 NTD structure_element Based on the titration curve for the chemical shift changes of L58, the apparent Kd between the isolated NTD and PIN was estimated to be 110 ± 5.8 μM. Considering the fact that the NTD and PIN domains are attached by a linker, the actual binding affinity is expected much higher in the native protein. RESULTS | |
| 113 116 PIN structure_element Based on the titration curve for the chemical shift changes of L58, the apparent Kd between the isolated NTD and PIN was estimated to be 110 ± 5.8 μM. Considering the fact that the NTD and PIN domains are attached by a linker, the actual binding affinity is expected much higher in the native protein. RESULTS | |
| 181 184 NTD structure_element Based on the titration curve for the chemical shift changes of L58, the apparent Kd between the isolated NTD and PIN was estimated to be 110 ± 5.8 μM. Considering the fact that the NTD and PIN domains are attached by a linker, the actual binding affinity is expected much higher in the native protein. RESULTS | |
| 189 192 PIN structure_element Based on the titration curve for the chemical shift changes of L58, the apparent Kd between the isolated NTD and PIN was estimated to be 110 ± 5.8 μM. Considering the fact that the NTD and PIN domains are attached by a linker, the actual binding affinity is expected much higher in the native protein. RESULTS | |
| 219 225 linker structure_element Based on the titration curve for the chemical shift changes of L58, the apparent Kd between the isolated NTD and PIN was estimated to be 110 ± 5.8 μM. Considering the fact that the NTD and PIN domains are attached by a linker, the actual binding affinity is expected much higher in the native protein. RESULTS | |
| 238 254 binding affinity evidence Based on the titration curve for the chemical shift changes of L58, the apparent Kd between the isolated NTD and PIN was estimated to be 110 ± 5.8 μM. Considering the fact that the NTD and PIN domains are attached by a linker, the actual binding affinity is expected much higher in the native protein. RESULTS | |
| 286 292 native protein_state Based on the titration curve for the chemical shift changes of L58, the apparent Kd between the isolated NTD and PIN was estimated to be 110 ± 5.8 μM. Considering the fact that the NTD and PIN domains are attached by a linker, the actual binding affinity is expected much higher in the native protein. RESULTS | |
| 70 87 PIN/NTD interface site Mapping the residues with chemical shift changes reveals the putative PIN/NTD interface, which includes a helix that harbors catalytic residues D225 and D226 on the PIN domain (Fig. 3a). RESULTS | |
| 106 111 helix structure_element Mapping the residues with chemical shift changes reveals the putative PIN/NTD interface, which includes a helix that harbors catalytic residues D225 and D226 on the PIN domain (Fig. 3a). RESULTS | |
| 144 148 D225 residue_name_number Mapping the residues with chemical shift changes reveals the putative PIN/NTD interface, which includes a helix that harbors catalytic residues D225 and D226 on the PIN domain (Fig. 3a). RESULTS | |
| 153 157 D226 residue_name_number Mapping the residues with chemical shift changes reveals the putative PIN/NTD interface, which includes a helix that harbors catalytic residues D225 and D226 on the PIN domain (Fig. 3a). RESULTS | |
| 165 168 PIN structure_element Mapping the residues with chemical shift changes reveals the putative PIN/NTD interface, which includes a helix that harbors catalytic residues D225 and D226 on the PIN domain (Fig. 3a). RESULTS | |
| 28 40 binding site site Interestingly, the putative binding site for the NTD overlaps with the PIN-PIN dimer interface, implying that NTD binding can “terminate” PIN-PIN oligomerization (Fig. 2b). RESULTS | |
| 49 52 NTD structure_element Interestingly, the putative binding site for the NTD overlaps with the PIN-PIN dimer interface, implying that NTD binding can “terminate” PIN-PIN oligomerization (Fig. 2b). RESULTS | |
| 71 94 PIN-PIN dimer interface site Interestingly, the putative binding site for the NTD overlaps with the PIN-PIN dimer interface, implying that NTD binding can “terminate” PIN-PIN oligomerization (Fig. 2b). RESULTS | |
| 110 113 NTD structure_element Interestingly, the putative binding site for the NTD overlaps with the PIN-PIN dimer interface, implying that NTD binding can “terminate” PIN-PIN oligomerization (Fig. 2b). RESULTS | |
| 138 141 PIN structure_element Interestingly, the putative binding site for the NTD overlaps with the PIN-PIN dimer interface, implying that NTD binding can “terminate” PIN-PIN oligomerization (Fig. 2b). RESULTS | |
| 142 145 PIN structure_element Interestingly, the putative binding site for the NTD overlaps with the PIN-PIN dimer interface, implying that NTD binding can “terminate” PIN-PIN oligomerization (Fig. 2b). RESULTS | |
| 3 20 in silico docking experimental_method An in silico docking of the NTD and PIN domains using chemical shift restraints provided a model consistent with the NMR experiments (Fig. 3c). RESULTS | |
| 28 31 NTD structure_element An in silico docking of the NTD and PIN domains using chemical shift restraints provided a model consistent with the NMR experiments (Fig. 3c). RESULTS | |
| 36 39 PIN structure_element An in silico docking of the NTD and PIN domains using chemical shift restraints provided a model consistent with the NMR experiments (Fig. 3c). RESULTS | |
| 54 79 chemical shift restraints evidence An in silico docking of the NTD and PIN domains using chemical shift restraints provided a model consistent with the NMR experiments (Fig. 3c). RESULTS | |
| 117 120 NMR experimental_method An in silico docking of the NTD and PIN domains using chemical shift restraints provided a model consistent with the NMR experiments (Fig. 3c). RESULTS | |
| 22 31 Regnase-1 protein Residues critical for Regnase-1 RNase activity RESULTS | |
| 32 37 RNase protein_type Residues critical for Regnase-1 RNase activity RESULTS | |
| 47 56 Regnase-1 protein To gain insight into the residues critical for Regnase-1 RNase activity, each basic or aromatic residue located around the catalytic site of the PIN oligomer was mutated to alanine, and the oligomerization and RNase activity were investigated (Fig. 4). RESULTS | |
| 57 62 RNase protein_type To gain insight into the residues critical for Regnase-1 RNase activity, each basic or aromatic residue located around the catalytic site of the PIN oligomer was mutated to alanine, and the oligomerization and RNase activity were investigated (Fig. 4). RESULTS | |
| 123 137 catalytic site site To gain insight into the residues critical for Regnase-1 RNase activity, each basic or aromatic residue located around the catalytic site of the PIN oligomer was mutated to alanine, and the oligomerization and RNase activity were investigated (Fig. 4). RESULTS | |
| 145 148 PIN structure_element To gain insight into the residues critical for Regnase-1 RNase activity, each basic or aromatic residue located around the catalytic site of the PIN oligomer was mutated to alanine, and the oligomerization and RNase activity were investigated (Fig. 4). RESULTS | |
| 149 157 oligomer oligomeric_state To gain insight into the residues critical for Regnase-1 RNase activity, each basic or aromatic residue located around the catalytic site of the PIN oligomer was mutated to alanine, and the oligomerization and RNase activity were investigated (Fig. 4). RESULTS | |
| 162 172 mutated to experimental_method To gain insight into the residues critical for Regnase-1 RNase activity, each basic or aromatic residue located around the catalytic site of the PIN oligomer was mutated to alanine, and the oligomerization and RNase activity were investigated (Fig. 4). RESULTS | |
| 173 180 alanine residue_name To gain insight into the residues critical for Regnase-1 RNase activity, each basic or aromatic residue located around the catalytic site of the PIN oligomer was mutated to alanine, and the oligomerization and RNase activity were investigated (Fig. 4). RESULTS | |
| 210 215 RNase protein_type To gain insight into the residues critical for Regnase-1 RNase activity, each basic or aromatic residue located around the catalytic site of the PIN oligomer was mutated to alanine, and the oligomerization and RNase activity were investigated (Fig. 4). RESULTS | |
| 9 30 gel filtration assays experimental_method From the gel filtration assays, all mutants except R214A formed dimers, suggesting that any lack of RNase activity in the mutants, except R214A, was directly due to mutational effects of the specific residues and not to abrogation of dimer formation. RESULTS | |
| 36 43 mutants protein_state From the gel filtration assays, all mutants except R214A formed dimers, suggesting that any lack of RNase activity in the mutants, except R214A, was directly due to mutational effects of the specific residues and not to abrogation of dimer formation. RESULTS | |
| 51 56 R214A mutant From the gel filtration assays, all mutants except R214A formed dimers, suggesting that any lack of RNase activity in the mutants, except R214A, was directly due to mutational effects of the specific residues and not to abrogation of dimer formation. RESULTS | |
| 64 70 dimers oligomeric_state From the gel filtration assays, all mutants except R214A formed dimers, suggesting that any lack of RNase activity in the mutants, except R214A, was directly due to mutational effects of the specific residues and not to abrogation of dimer formation. RESULTS | |
| 100 105 RNase protein_type From the gel filtration assays, all mutants except R214A formed dimers, suggesting that any lack of RNase activity in the mutants, except R214A, was directly due to mutational effects of the specific residues and not to abrogation of dimer formation. RESULTS | |
| 122 129 mutants protein_state From the gel filtration assays, all mutants except R214A formed dimers, suggesting that any lack of RNase activity in the mutants, except R214A, was directly due to mutational effects of the specific residues and not to abrogation of dimer formation. RESULTS | |
| 138 143 R214A mutant From the gel filtration assays, all mutants except R214A formed dimers, suggesting that any lack of RNase activity in the mutants, except R214A, was directly due to mutational effects of the specific residues and not to abrogation of dimer formation. RESULTS | |
| 234 239 dimer oligomeric_state From the gel filtration assays, all mutants except R214A formed dimers, suggesting that any lack of RNase activity in the mutants, except R214A, was directly due to mutational effects of the specific residues and not to abrogation of dimer formation. RESULTS | |
| 4 9 W182A mutant The W182A, R183A, and R214A mutants markedly lost cleavage activity for IL-6 mRNA as well as for Regnase-1 mRNA. RESULTS | |
| 11 16 R183A mutant The W182A, R183A, and R214A mutants markedly lost cleavage activity for IL-6 mRNA as well as for Regnase-1 mRNA. RESULTS | |
| 22 27 R214A mutant The W182A, R183A, and R214A mutants markedly lost cleavage activity for IL-6 mRNA as well as for Regnase-1 mRNA. RESULTS | |
| 28 35 mutants protein_state The W182A, R183A, and R214A mutants markedly lost cleavage activity for IL-6 mRNA as well as for Regnase-1 mRNA. RESULTS | |
| 72 76 IL-6 protein_type The W182A, R183A, and R214A mutants markedly lost cleavage activity for IL-6 mRNA as well as for Regnase-1 mRNA. RESULTS | |
| 77 81 mRNA chemical The W182A, R183A, and R214A mutants markedly lost cleavage activity for IL-6 mRNA as well as for Regnase-1 mRNA. RESULTS | |
| 97 106 Regnase-1 protein The W182A, R183A, and R214A mutants markedly lost cleavage activity for IL-6 mRNA as well as for Regnase-1 mRNA. RESULTS | |
| 107 111 mRNA chemical The W182A, R183A, and R214A mutants markedly lost cleavage activity for IL-6 mRNA as well as for Regnase-1 mRNA. RESULTS | |
| 4 9 K184A mutant The K184A, R215A, and R220A mutants moderately but significantly decreased the cleavage activity for both target mRNAs. RESULTS | |
| 11 16 R215A mutant The K184A, R215A, and R220A mutants moderately but significantly decreased the cleavage activity for both target mRNAs. RESULTS | |
| 22 27 R220A mutant The K184A, R215A, and R220A mutants moderately but significantly decreased the cleavage activity for both target mRNAs. RESULTS | |
| 28 35 mutants protein_state The K184A, R215A, and R220A mutants moderately but significantly decreased the cleavage activity for both target mRNAs. RESULTS | |
| 113 118 mRNAs chemical The K184A, R215A, and R220A mutants moderately but significantly decreased the cleavage activity for both target mRNAs. RESULTS | |
| 18 22 K219 residue_name_number The importance of K219 and R247 was slightly different for IL-6 and Regnase-1 mRNA; both K219 and R247 were more important in the cleavage of IL-6 mRNA than for Regnase-1 mRNA. RESULTS | |
| 27 31 R247 residue_name_number The importance of K219 and R247 was slightly different for IL-6 and Regnase-1 mRNA; both K219 and R247 were more important in the cleavage of IL-6 mRNA than for Regnase-1 mRNA. RESULTS | |
| 59 63 IL-6 protein_type The importance of K219 and R247 was slightly different for IL-6 and Regnase-1 mRNA; both K219 and R247 were more important in the cleavage of IL-6 mRNA than for Regnase-1 mRNA. RESULTS | |
| 68 77 Regnase-1 protein The importance of K219 and R247 was slightly different for IL-6 and Regnase-1 mRNA; both K219 and R247 were more important in the cleavage of IL-6 mRNA than for Regnase-1 mRNA. RESULTS | |
| 78 82 mRNA chemical The importance of K219 and R247 was slightly different for IL-6 and Regnase-1 mRNA; both K219 and R247 were more important in the cleavage of IL-6 mRNA than for Regnase-1 mRNA. RESULTS | |
| 89 93 K219 residue_name_number The importance of K219 and R247 was slightly different for IL-6 and Regnase-1 mRNA; both K219 and R247 were more important in the cleavage of IL-6 mRNA than for Regnase-1 mRNA. RESULTS | |
| 98 102 R247 residue_name_number The importance of K219 and R247 was slightly different for IL-6 and Regnase-1 mRNA; both K219 and R247 were more important in the cleavage of IL-6 mRNA than for Regnase-1 mRNA. RESULTS | |
| 142 146 IL-6 protein_type The importance of K219 and R247 was slightly different for IL-6 and Regnase-1 mRNA; both K219 and R247 were more important in the cleavage of IL-6 mRNA than for Regnase-1 mRNA. RESULTS | |
| 147 151 mRNA chemical The importance of K219 and R247 was slightly different for IL-6 and Regnase-1 mRNA; both K219 and R247 were more important in the cleavage of IL-6 mRNA than for Regnase-1 mRNA. RESULTS | |
| 161 170 Regnase-1 protein The importance of K219 and R247 was slightly different for IL-6 and Regnase-1 mRNA; both K219 and R247 were more important in the cleavage of IL-6 mRNA than for Regnase-1 mRNA. RESULTS | |
| 171 175 mRNA chemical The importance of K219 and R247 was slightly different for IL-6 and Regnase-1 mRNA; both K219 and R247 were more important in the cleavage of IL-6 mRNA than for Regnase-1 mRNA. RESULTS | |
| 27 31 K152 residue_name_number The other mutated residues—K152, R158, R188, R200, K204, K206, K257, and R258—were not critical for RNase activity. RESULTS | |
| 33 37 R158 residue_name_number The other mutated residues—K152, R158, R188, R200, K204, K206, K257, and R258—were not critical for RNase activity. RESULTS | |
| 39 43 R188 residue_name_number The other mutated residues—K152, R158, R188, R200, K204, K206, K257, and R258—were not critical for RNase activity. RESULTS | |
| 45 49 R200 residue_name_number The other mutated residues—K152, R158, R188, R200, K204, K206, K257, and R258—were not critical for RNase activity. RESULTS | |
| 51 55 K204 residue_name_number The other mutated residues—K152, R158, R188, R200, K204, K206, K257, and R258—were not critical for RNase activity. RESULTS | |
| 57 61 K206 residue_name_number The other mutated residues—K152, R158, R188, R200, K204, K206, K257, and R258—were not critical for RNase activity. RESULTS | |
| 63 67 K257 residue_name_number The other mutated residues—K152, R158, R188, R200, K204, K206, K257, and R258—were not critical for RNase activity. RESULTS | |
| 73 77 R258 residue_name_number The other mutated residues—K152, R158, R188, R200, K204, K206, K257, and R258—were not critical for RNase activity. RESULTS | |
| 100 105 RNase protein_type The other mutated residues—K152, R158, R188, R200, K204, K206, K257, and R258—were not critical for RNase activity. RESULTS | |
| 27 31 W182 residue_name_number The importance of residues W182 and R183 can readily be understood in terms of the monomeric PIN structure as they are located near to the RNase catalytic site; however, the importance of residue K184, which points away from the active site is more easily rationalized in terms of the oligomeric structure, in which the “secondary” chain’s residue K184 is positioned near the “primary” chain’s catalytic site (Fig. 4). RESULTS | |
| 36 40 R183 residue_name_number The importance of residues W182 and R183 can readily be understood in terms of the monomeric PIN structure as they are located near to the RNase catalytic site; however, the importance of residue K184, which points away from the active site is more easily rationalized in terms of the oligomeric structure, in which the “secondary” chain’s residue K184 is positioned near the “primary” chain’s catalytic site (Fig. 4). RESULTS | |
| 83 92 monomeric oligomeric_state The importance of residues W182 and R183 can readily be understood in terms of the monomeric PIN structure as they are located near to the RNase catalytic site; however, the importance of residue K184, which points away from the active site is more easily rationalized in terms of the oligomeric structure, in which the “secondary” chain’s residue K184 is positioned near the “primary” chain’s catalytic site (Fig. 4). RESULTS | |
| 93 96 PIN structure_element The importance of residues W182 and R183 can readily be understood in terms of the monomeric PIN structure as they are located near to the RNase catalytic site; however, the importance of residue K184, which points away from the active site is more easily rationalized in terms of the oligomeric structure, in which the “secondary” chain’s residue K184 is positioned near the “primary” chain’s catalytic site (Fig. 4). RESULTS | |
| 97 106 structure evidence The importance of residues W182 and R183 can readily be understood in terms of the monomeric PIN structure as they are located near to the RNase catalytic site; however, the importance of residue K184, which points away from the active site is more easily rationalized in terms of the oligomeric structure, in which the “secondary” chain’s residue K184 is positioned near the “primary” chain’s catalytic site (Fig. 4). RESULTS | |
| 139 144 RNase protein_type The importance of residues W182 and R183 can readily be understood in terms of the monomeric PIN structure as they are located near to the RNase catalytic site; however, the importance of residue K184, which points away from the active site is more easily rationalized in terms of the oligomeric structure, in which the “secondary” chain’s residue K184 is positioned near the “primary” chain’s catalytic site (Fig. 4). RESULTS | |
| 145 159 catalytic site site The importance of residues W182 and R183 can readily be understood in terms of the monomeric PIN structure as they are located near to the RNase catalytic site; however, the importance of residue K184, which points away from the active site is more easily rationalized in terms of the oligomeric structure, in which the “secondary” chain’s residue K184 is positioned near the “primary” chain’s catalytic site (Fig. 4). RESULTS | |
| 196 200 K184 residue_name_number The importance of residues W182 and R183 can readily be understood in terms of the monomeric PIN structure as they are located near to the RNase catalytic site; however, the importance of residue K184, which points away from the active site is more easily rationalized in terms of the oligomeric structure, in which the “secondary” chain’s residue K184 is positioned near the “primary” chain’s catalytic site (Fig. 4). RESULTS | |
| 229 240 active site site The importance of residues W182 and R183 can readily be understood in terms of the monomeric PIN structure as they are located near to the RNase catalytic site; however, the importance of residue K184, which points away from the active site is more easily rationalized in terms of the oligomeric structure, in which the “secondary” chain’s residue K184 is positioned near the “primary” chain’s catalytic site (Fig. 4). RESULTS | |
| 296 305 structure evidence The importance of residues W182 and R183 can readily be understood in terms of the monomeric PIN structure as they are located near to the RNase catalytic site; however, the importance of residue K184, which points away from the active site is more easily rationalized in terms of the oligomeric structure, in which the “secondary” chain’s residue K184 is positioned near the “primary” chain’s catalytic site (Fig. 4). RESULTS | |
| 348 352 K184 residue_name_number The importance of residues W182 and R183 can readily be understood in terms of the monomeric PIN structure as they are located near to the RNase catalytic site; however, the importance of residue K184, which points away from the active site is more easily rationalized in terms of the oligomeric structure, in which the “secondary” chain’s residue K184 is positioned near the “primary” chain’s catalytic site (Fig. 4). RESULTS | |
| 377 385 primary” protein_state The importance of residues W182 and R183 can readily be understood in terms of the monomeric PIN structure as they are located near to the RNase catalytic site; however, the importance of residue K184, which points away from the active site is more easily rationalized in terms of the oligomeric structure, in which the “secondary” chain’s residue K184 is positioned near the “primary” chain’s catalytic site (Fig. 4). RESULTS | |
| 394 408 catalytic site site The importance of residues W182 and R183 can readily be understood in terms of the monomeric PIN structure as they are located near to the RNase catalytic site; however, the importance of residue K184, which points away from the active site is more easily rationalized in terms of the oligomeric structure, in which the “secondary” chain’s residue K184 is positioned near the “primary” chain’s catalytic site (Fig. 4). RESULTS | |
| 13 17 R214 residue_name_number In contrast, R214 is important for oligomerization of the PIN domain and the “secondary” chain’s residue R214 is also positioned near the “primary” chain’s active site within the dimer interface. RESULTS | |
| 58 61 PIN structure_element In contrast, R214 is important for oligomerization of the PIN domain and the “secondary” chain’s residue R214 is also positioned near the “primary” chain’s active site within the dimer interface. RESULTS | |
| 105 109 R214 residue_name_number In contrast, R214 is important for oligomerization of the PIN domain and the “secondary” chain’s residue R214 is also positioned near the “primary” chain’s active site within the dimer interface. RESULTS | |
| 139 146 primary protein_state In contrast, R214 is important for oligomerization of the PIN domain and the “secondary” chain’s residue R214 is also positioned near the “primary” chain’s active site within the dimer interface. RESULTS | |
| 156 167 active site site In contrast, R214 is important for oligomerization of the PIN domain and the “secondary” chain’s residue R214 is also positioned near the “primary” chain’s active site within the dimer interface. RESULTS | |
| 179 194 dimer interface site In contrast, R214 is important for oligomerization of the PIN domain and the “secondary” chain’s residue R214 is also positioned near the “primary” chain’s active site within the dimer interface. RESULTS | |
| 28 57 putative-RNA binding residues site It should be noted that the putative-RNA binding residues K184 and R214 are unique to Regnase-1 among PIN domains. RESULTS | |
| 58 62 K184 residue_name_number It should be noted that the putative-RNA binding residues K184 and R214 are unique to Regnase-1 among PIN domains. RESULTS | |
| 67 71 R214 residue_name_number It should be noted that the putative-RNA binding residues K184 and R214 are unique to Regnase-1 among PIN domains. RESULTS | |
| 86 95 Regnase-1 protein It should be noted that the putative-RNA binding residues K184 and R214 are unique to Regnase-1 among PIN domains. RESULTS | |
| 102 105 PIN structure_element It should be noted that the putative-RNA binding residues K184 and R214 are unique to Regnase-1 among PIN domains. RESULTS | |
| 30 34 mRNA chemical Molecular mechanism of target mRNA cleavage by the PIN dimer RESULTS | |
| 51 54 PIN structure_element Molecular mechanism of target mRNA cleavage by the PIN dimer RESULTS | |
| 55 60 dimer oligomeric_state Molecular mechanism of target mRNA cleavage by the PIN dimer RESULTS | |
| 4 26 mutational experiments experimental_method Our mutational experiments indicated that the observed dimer is functional and that the role of the secondary PIN domain is to position Regnase-1-unique RNA binding residues near the active site of the primary PIN domain. RESULTS | |
| 55 60 dimer oligomeric_state Our mutational experiments indicated that the observed dimer is functional and that the role of the secondary PIN domain is to position Regnase-1-unique RNA binding residues near the active site of the primary PIN domain. RESULTS | |
| 100 109 secondary protein_state Our mutational experiments indicated that the observed dimer is functional and that the role of the secondary PIN domain is to position Regnase-1-unique RNA binding residues near the active site of the primary PIN domain. RESULTS | |
| 110 113 PIN structure_element Our mutational experiments indicated that the observed dimer is functional and that the role of the secondary PIN domain is to position Regnase-1-unique RNA binding residues near the active site of the primary PIN domain. RESULTS | |
| 136 145 Regnase-1 protein Our mutational experiments indicated that the observed dimer is functional and that the role of the secondary PIN domain is to position Regnase-1-unique RNA binding residues near the active site of the primary PIN domain. RESULTS | |
| 153 173 RNA binding residues site Our mutational experiments indicated that the observed dimer is functional and that the role of the secondary PIN domain is to position Regnase-1-unique RNA binding residues near the active site of the primary PIN domain. RESULTS | |
| 183 194 active site site Our mutational experiments indicated that the observed dimer is functional and that the role of the secondary PIN domain is to position Regnase-1-unique RNA binding residues near the active site of the primary PIN domain. RESULTS | |
| 202 209 primary protein_state Our mutational experiments indicated that the observed dimer is functional and that the role of the secondary PIN domain is to position Regnase-1-unique RNA binding residues near the active site of the primary PIN domain. RESULTS | |
| 210 213 PIN structure_element Our mutational experiments indicated that the observed dimer is functional and that the role of the secondary PIN domain is to position Regnase-1-unique RNA binding residues near the active site of the primary PIN domain. RESULTS | |
| 50 72 catalytically inactive protein_state If this model is correct, then we reasoned that a catalytically inactive PIN and a PIN lacking the putative RNA-binding residues ought to be inactive in isolation but become active when mixed together. RESULTS | |
| 73 76 PIN structure_element If this model is correct, then we reasoned that a catalytically inactive PIN and a PIN lacking the putative RNA-binding residues ought to be inactive in isolation but become active when mixed together. RESULTS | |
| 83 86 PIN structure_element If this model is correct, then we reasoned that a catalytically inactive PIN and a PIN lacking the putative RNA-binding residues ought to be inactive in isolation but become active when mixed together. RESULTS | |
| 87 94 lacking protein_state If this model is correct, then we reasoned that a catalytically inactive PIN and a PIN lacking the putative RNA-binding residues ought to be inactive in isolation but become active when mixed together. RESULTS | |
| 108 128 RNA-binding residues site If this model is correct, then we reasoned that a catalytically inactive PIN and a PIN lacking the putative RNA-binding residues ought to be inactive in isolation but become active when mixed together. RESULTS | |
| 141 149 inactive protein_state If this model is correct, then we reasoned that a catalytically inactive PIN and a PIN lacking the putative RNA-binding residues ought to be inactive in isolation but become active when mixed together. RESULTS | |
| 174 180 active protein_state If this model is correct, then we reasoned that a catalytically inactive PIN and a PIN lacking the putative RNA-binding residues ought to be inactive in isolation but become active when mixed together. RESULTS | |
| 47 71 in vitro cleavage assays experimental_method In order to test this hypothesis, we performed in vitro cleavage assays using combinations of Regnase-1 mutants that had no or decreased RNase activities by themselves (Fig. 5). RESULTS | |
| 94 103 Regnase-1 protein In order to test this hypothesis, we performed in vitro cleavage assays using combinations of Regnase-1 mutants that had no or decreased RNase activities by themselves (Fig. 5). RESULTS | |
| 104 111 mutants protein_state In order to test this hypothesis, we performed in vitro cleavage assays using combinations of Regnase-1 mutants that had no or decreased RNase activities by themselves (Fig. 5). RESULTS | |
| 137 142 RNase protein_type In order to test this hypothesis, we performed in vitro cleavage assays using combinations of Regnase-1 mutants that had no or decreased RNase activities by themselves (Fig. 5). RESULTS | |
| 23 43 catalytically active protein_state One group consisted of catalytically active PIN domains with mutation of basic residues found in the previous section to confer decreased RNase activity (Fig. 4). RESULTS | |
| 44 47 PIN structure_element One group consisted of catalytically active PIN domains with mutation of basic residues found in the previous section to confer decreased RNase activity (Fig. 4). RESULTS | |
| 61 72 mutation of experimental_method One group consisted of catalytically active PIN domains with mutation of basic residues found in the previous section to confer decreased RNase activity (Fig. 4). RESULTS | |
| 138 143 RNase protein_type One group consisted of catalytically active PIN domains with mutation of basic residues found in the previous section to confer decreased RNase activity (Fig. 4). RESULTS | |
| 25 29 DDNN mutant These were paired with a DDNN mutant that had no RNase activity by itself. RESULTS | |
| 30 36 mutant protein_state These were paired with a DDNN mutant that had no RNase activity by itself. RESULTS | |
| 49 54 RNase protein_type These were paired with a DDNN mutant that had no RNase activity by itself. RESULTS | |
| 59 71 heterodimers oligomeric_state When any members of the two groups are mixed, two kinds of heterodimers can be formed: one is composed of a DDNN primary PIN and a basic residue mutant secondary PIN and is expected to exhibit no RNase activity; the other is composed of a basic residue mutant primary PIN and a DDNN secondary PIN and is predicted to rescue RNase activity (Fig. 5a). RESULTS | |
| 108 112 DDNN mutant When any members of the two groups are mixed, two kinds of heterodimers can be formed: one is composed of a DDNN primary PIN and a basic residue mutant secondary PIN and is expected to exhibit no RNase activity; the other is composed of a basic residue mutant primary PIN and a DDNN secondary PIN and is predicted to rescue RNase activity (Fig. 5a). RESULTS | |
| 113 120 primary protein_state When any members of the two groups are mixed, two kinds of heterodimers can be formed: one is composed of a DDNN primary PIN and a basic residue mutant secondary PIN and is expected to exhibit no RNase activity; the other is composed of a basic residue mutant primary PIN and a DDNN secondary PIN and is predicted to rescue RNase activity (Fig. 5a). RESULTS | |
| 121 124 PIN structure_element When any members of the two groups are mixed, two kinds of heterodimers can be formed: one is composed of a DDNN primary PIN and a basic residue mutant secondary PIN and is expected to exhibit no RNase activity; the other is composed of a basic residue mutant primary PIN and a DDNN secondary PIN and is predicted to rescue RNase activity (Fig. 5a). RESULTS | |
| 145 151 mutant protein_state When any members of the two groups are mixed, two kinds of heterodimers can be formed: one is composed of a DDNN primary PIN and a basic residue mutant secondary PIN and is expected to exhibit no RNase activity; the other is composed of a basic residue mutant primary PIN and a DDNN secondary PIN and is predicted to rescue RNase activity (Fig. 5a). RESULTS | |
| 152 161 secondary protein_state When any members of the two groups are mixed, two kinds of heterodimers can be formed: one is composed of a DDNN primary PIN and a basic residue mutant secondary PIN and is expected to exhibit no RNase activity; the other is composed of a basic residue mutant primary PIN and a DDNN secondary PIN and is predicted to rescue RNase activity (Fig. 5a). RESULTS | |
| 162 165 PIN structure_element When any members of the two groups are mixed, two kinds of heterodimers can be formed: one is composed of a DDNN primary PIN and a basic residue mutant secondary PIN and is expected to exhibit no RNase activity; the other is composed of a basic residue mutant primary PIN and a DDNN secondary PIN and is predicted to rescue RNase activity (Fig. 5a). RESULTS | |
| 196 201 RNase protein_type When any members of the two groups are mixed, two kinds of heterodimers can be formed: one is composed of a DDNN primary PIN and a basic residue mutant secondary PIN and is expected to exhibit no RNase activity; the other is composed of a basic residue mutant primary PIN and a DDNN secondary PIN and is predicted to rescue RNase activity (Fig. 5a). RESULTS | |
| 253 259 mutant protein_state When any members of the two groups are mixed, two kinds of heterodimers can be formed: one is composed of a DDNN primary PIN and a basic residue mutant secondary PIN and is expected to exhibit no RNase activity; the other is composed of a basic residue mutant primary PIN and a DDNN secondary PIN and is predicted to rescue RNase activity (Fig. 5a). RESULTS | |
| 260 267 primary protein_state When any members of the two groups are mixed, two kinds of heterodimers can be formed: one is composed of a DDNN primary PIN and a basic residue mutant secondary PIN and is expected to exhibit no RNase activity; the other is composed of a basic residue mutant primary PIN and a DDNN secondary PIN and is predicted to rescue RNase activity (Fig. 5a). RESULTS | |
| 268 271 PIN structure_element When any members of the two groups are mixed, two kinds of heterodimers can be formed: one is composed of a DDNN primary PIN and a basic residue mutant secondary PIN and is expected to exhibit no RNase activity; the other is composed of a basic residue mutant primary PIN and a DDNN secondary PIN and is predicted to rescue RNase activity (Fig. 5a). RESULTS | |
| 278 282 DDNN mutant When any members of the two groups are mixed, two kinds of heterodimers can be formed: one is composed of a DDNN primary PIN and a basic residue mutant secondary PIN and is expected to exhibit no RNase activity; the other is composed of a basic residue mutant primary PIN and a DDNN secondary PIN and is predicted to rescue RNase activity (Fig. 5a). RESULTS | |
| 283 292 secondary protein_state When any members of the two groups are mixed, two kinds of heterodimers can be formed: one is composed of a DDNN primary PIN and a basic residue mutant secondary PIN and is expected to exhibit no RNase activity; the other is composed of a basic residue mutant primary PIN and a DDNN secondary PIN and is predicted to rescue RNase activity (Fig. 5a). RESULTS | |
| 293 296 PIN structure_element When any members of the two groups are mixed, two kinds of heterodimers can be formed: one is composed of a DDNN primary PIN and a basic residue mutant secondary PIN and is expected to exhibit no RNase activity; the other is composed of a basic residue mutant primary PIN and a DDNN secondary PIN and is predicted to rescue RNase activity (Fig. 5a). RESULTS | |
| 324 329 RNase protein_type When any members of the two groups are mixed, two kinds of heterodimers can be formed: one is composed of a DDNN primary PIN and a basic residue mutant secondary PIN and is expected to exhibit no RNase activity; the other is composed of a basic residue mutant primary PIN and a DDNN secondary PIN and is predicted to rescue RNase activity (Fig. 5a). RESULTS | |
| 21 43 fluorescence intensity evidence When we compared the fluorescence intensity of uncleaved IL-6 mRNA, basic residue mutants W182A, K184A, R214A, and R220A were rescued upon addition of the DDNN mutant (Fig. 5b). RESULTS | |
| 47 56 uncleaved protein_state When we compared the fluorescence intensity of uncleaved IL-6 mRNA, basic residue mutants W182A, K184A, R214A, and R220A were rescued upon addition of the DDNN mutant (Fig. 5b). RESULTS | |
| 57 61 IL-6 protein_type When we compared the fluorescence intensity of uncleaved IL-6 mRNA, basic residue mutants W182A, K184A, R214A, and R220A were rescued upon addition of the DDNN mutant (Fig. 5b). RESULTS | |
| 62 66 mRNA chemical When we compared the fluorescence intensity of uncleaved IL-6 mRNA, basic residue mutants W182A, K184A, R214A, and R220A were rescued upon addition of the DDNN mutant (Fig. 5b). RESULTS | |
| 82 89 mutants protein_state When we compared the fluorescence intensity of uncleaved IL-6 mRNA, basic residue mutants W182A, K184A, R214A, and R220A were rescued upon addition of the DDNN mutant (Fig. 5b). RESULTS | |
| 90 95 W182A mutant When we compared the fluorescence intensity of uncleaved IL-6 mRNA, basic residue mutants W182A, K184A, R214A, and R220A were rescued upon addition of the DDNN mutant (Fig. 5b). RESULTS | |
| 97 102 K184A mutant When we compared the fluorescence intensity of uncleaved IL-6 mRNA, basic residue mutants W182A, K184A, R214A, and R220A were rescued upon addition of the DDNN mutant (Fig. 5b). RESULTS | |
| 104 109 R214A mutant When we compared the fluorescence intensity of uncleaved IL-6 mRNA, basic residue mutants W182A, K184A, R214A, and R220A were rescued upon addition of the DDNN mutant (Fig. 5b). RESULTS | |
| 115 120 R220A mutant When we compared the fluorescence intensity of uncleaved IL-6 mRNA, basic residue mutants W182A, K184A, R214A, and R220A were rescued upon addition of the DDNN mutant (Fig. 5b). RESULTS | |
| 155 159 DDNN mutant When we compared the fluorescence intensity of uncleaved IL-6 mRNA, basic residue mutants W182A, K184A, R214A, and R220A were rescued upon addition of the DDNN mutant (Fig. 5b). RESULTS | |
| 160 166 mutant protein_state When we compared the fluorescence intensity of uncleaved IL-6 mRNA, basic residue mutants W182A, K184A, R214A, and R220A were rescued upon addition of the DDNN mutant (Fig. 5b). RESULTS | |
| 35 57 fluorescence intensity evidence Consistently, when we compared the fluorescence intensity of the uncleaved Regnase-1 mRNA, basic residue mutants K184A and R214A were rescued upon addition of the DDNN mutant (Fig. 5c). RESULTS | |
| 65 74 uncleaved protein_state Consistently, when we compared the fluorescence intensity of the uncleaved Regnase-1 mRNA, basic residue mutants K184A and R214A were rescued upon addition of the DDNN mutant (Fig. 5c). RESULTS | |
| 75 84 Regnase-1 protein Consistently, when we compared the fluorescence intensity of the uncleaved Regnase-1 mRNA, basic residue mutants K184A and R214A were rescued upon addition of the DDNN mutant (Fig. 5c). RESULTS | |
| 85 89 mRNA chemical Consistently, when we compared the fluorescence intensity of the uncleaved Regnase-1 mRNA, basic residue mutants K184A and R214A were rescued upon addition of the DDNN mutant (Fig. 5c). RESULTS | |
| 105 112 mutants protein_state Consistently, when we compared the fluorescence intensity of the uncleaved Regnase-1 mRNA, basic residue mutants K184A and R214A were rescued upon addition of the DDNN mutant (Fig. 5c). RESULTS | |
| 113 118 K184A mutant Consistently, when we compared the fluorescence intensity of the uncleaved Regnase-1 mRNA, basic residue mutants K184A and R214A were rescued upon addition of the DDNN mutant (Fig. 5c). RESULTS | |
| 123 128 R214A mutant Consistently, when we compared the fluorescence intensity of the uncleaved Regnase-1 mRNA, basic residue mutants K184A and R214A were rescued upon addition of the DDNN mutant (Fig. 5c). RESULTS | |
| 163 167 DDNN mutant Consistently, when we compared the fluorescence intensity of the uncleaved Regnase-1 mRNA, basic residue mutants K184A and R214A were rescued upon addition of the DDNN mutant (Fig. 5c). RESULTS | |
| 168 174 mutant protein_state Consistently, when we compared the fluorescence intensity of the uncleaved Regnase-1 mRNA, basic residue mutants K184A and R214A were rescued upon addition of the DDNN mutant (Fig. 5c). RESULTS | |
| 10 15 K184A mutant Rescue of K184A and R214A by the DDNN mutant was also confirmed by a significant increase in the cleaved products. RESULTS | |
| 20 25 R214A mutant Rescue of K184A and R214A by the DDNN mutant was also confirmed by a significant increase in the cleaved products. RESULTS | |
| 33 37 DDNN mutant Rescue of K184A and R214A by the DDNN mutant was also confirmed by a significant increase in the cleaved products. RESULTS | |
| 38 44 mutant protein_state Rescue of K184A and R214A by the DDNN mutant was also confirmed by a significant increase in the cleaved products. RESULTS | |
| 60 64 K184 residue_name_number This is particularly significant because the side chains of K184 and R214 in the primary PIN are oriented away from their own catalytic center, while those in the secondary PIN face toward the catalytic center of the primary PIN. RESULTS | |
| 69 73 R214 residue_name_number This is particularly significant because the side chains of K184 and R214 in the primary PIN are oriented away from their own catalytic center, while those in the secondary PIN face toward the catalytic center of the primary PIN. RESULTS | |
| 81 88 primary protein_state This is particularly significant because the side chains of K184 and R214 in the primary PIN are oriented away from their own catalytic center, while those in the secondary PIN face toward the catalytic center of the primary PIN. RESULTS | |
| 89 92 PIN structure_element This is particularly significant because the side chains of K184 and R214 in the primary PIN are oriented away from their own catalytic center, while those in the secondary PIN face toward the catalytic center of the primary PIN. RESULTS | |
| 126 142 catalytic center site This is particularly significant because the side chains of K184 and R214 in the primary PIN are oriented away from their own catalytic center, while those in the secondary PIN face toward the catalytic center of the primary PIN. RESULTS | |
| 163 172 secondary protein_state This is particularly significant because the side chains of K184 and R214 in the primary PIN are oriented away from their own catalytic center, while those in the secondary PIN face toward the catalytic center of the primary PIN. RESULTS | |
| 173 176 PIN structure_element This is particularly significant because the side chains of K184 and R214 in the primary PIN are oriented away from their own catalytic center, while those in the secondary PIN face toward the catalytic center of the primary PIN. RESULTS | |
| 193 209 catalytic center site This is particularly significant because the side chains of K184 and R214 in the primary PIN are oriented away from their own catalytic center, while those in the secondary PIN face toward the catalytic center of the primary PIN. RESULTS | |
| 217 224 primary protein_state This is particularly significant because the side chains of K184 and R214 in the primary PIN are oriented away from their own catalytic center, while those in the secondary PIN face toward the catalytic center of the primary PIN. RESULTS | |
| 225 228 PIN structure_element This is particularly significant because the side chains of K184 and R214 in the primary PIN are oriented away from their own catalytic center, while those in the secondary PIN face toward the catalytic center of the primary PIN. RESULTS | |
| 0 4 R214 residue_name_number R214 is an important residue for dimer formation as shown in Fig. 2, therefore, R214A in the secondary PIN cannot dimerize. RESULTS | |
| 33 38 dimer oligomeric_state R214 is an important residue for dimer formation as shown in Fig. 2, therefore, R214A in the secondary PIN cannot dimerize. RESULTS | |
| 80 85 R214A mutant R214 is an important residue for dimer formation as shown in Fig. 2, therefore, R214A in the secondary PIN cannot dimerize. RESULTS | |
| 93 102 secondary protein_state R214 is an important residue for dimer formation as shown in Fig. 2, therefore, R214A in the secondary PIN cannot dimerize. RESULTS | |
| 103 106 PIN structure_element R214 is an important residue for dimer formation as shown in Fig. 2, therefore, R214A in the secondary PIN cannot dimerize. RESULTS | |
| 36 41 R214A mutant According to the proposed model, an R214A PIN domain can only form a dimer when the DDNN PIN acts as the secondary PIN. RESULTS | |
| 42 45 PIN structure_element According to the proposed model, an R214A PIN domain can only form a dimer when the DDNN PIN acts as the secondary PIN. RESULTS | |
| 69 74 dimer oligomeric_state According to the proposed model, an R214A PIN domain can only form a dimer when the DDNN PIN acts as the secondary PIN. RESULTS | |
| 84 88 DDNN mutant According to the proposed model, an R214A PIN domain can only form a dimer when the DDNN PIN acts as the secondary PIN. RESULTS | |
| 89 92 PIN structure_element According to the proposed model, an R214A PIN domain can only form a dimer when the DDNN PIN acts as the secondary PIN. RESULTS | |
| 105 114 secondary protein_state According to the proposed model, an R214A PIN domain can only form a dimer when the DDNN PIN acts as the secondary PIN. RESULTS | |
| 115 118 PIN structure_element According to the proposed model, an R214A PIN domain can only form a dimer when the DDNN PIN acts as the secondary PIN. RESULTS | |
| 85 97 head-to-tail protein_state Taken together, the rescue experiments above support the proposed model in which the head-to-tail dimer is functional in vitro. RESULTS | |
| 98 103 dimer oligomeric_state Taken together, the rescue experiments above support the proposed model in which the head-to-tail dimer is functional in vitro. RESULTS | |
| 36 46 structures evidence We determined the individual domain structures of Regnase-1 by NMR and X-ray crystallography. DISCUSS | |
| 50 59 Regnase-1 protein We determined the individual domain structures of Regnase-1 by NMR and X-ray crystallography. DISCUSS | |
| 63 66 NMR experimental_method We determined the individual domain structures of Regnase-1 by NMR and X-ray crystallography. DISCUSS | |
| 71 92 X-ray crystallography experimental_method We determined the individual domain structures of Regnase-1 by NMR and X-ray crystallography. DISCUSS | |
| 29 32 CTD structure_element Although the function of the CTD remains elusive, we revealed the functions of the NTD, PIN, and ZF domains. DISCUSS | |
| 83 86 NTD structure_element Although the function of the CTD remains elusive, we revealed the functions of the NTD, PIN, and ZF domains. DISCUSS | |
| 88 91 PIN structure_element Although the function of the CTD remains elusive, we revealed the functions of the NTD, PIN, and ZF domains. DISCUSS | |
| 97 99 ZF structure_element Although the function of the CTD remains elusive, we revealed the functions of the NTD, PIN, and ZF domains. DISCUSS | |
| 2 11 Regnase-1 protein A Regnase-1 construct consisting of PIN and ZF domains derived from Mus musculus was crystallized; however, the electron density of the ZF domain was low, indicating that the ZF domain is highly mobile in the absence of target mRNA or possibly other protein-protein interactions. DISCUSS | |
| 36 39 PIN structure_element A Regnase-1 construct consisting of PIN and ZF domains derived from Mus musculus was crystallized; however, the electron density of the ZF domain was low, indicating that the ZF domain is highly mobile in the absence of target mRNA or possibly other protein-protein interactions. DISCUSS | |
| 44 46 ZF structure_element A Regnase-1 construct consisting of PIN and ZF domains derived from Mus musculus was crystallized; however, the electron density of the ZF domain was low, indicating that the ZF domain is highly mobile in the absence of target mRNA or possibly other protein-protein interactions. DISCUSS | |
| 68 80 Mus musculus species A Regnase-1 construct consisting of PIN and ZF domains derived from Mus musculus was crystallized; however, the electron density of the ZF domain was low, indicating that the ZF domain is highly mobile in the absence of target mRNA or possibly other protein-protein interactions. DISCUSS | |
| 85 97 crystallized experimental_method A Regnase-1 construct consisting of PIN and ZF domains derived from Mus musculus was crystallized; however, the electron density of the ZF domain was low, indicating that the ZF domain is highly mobile in the absence of target mRNA or possibly other protein-protein interactions. DISCUSS | |
| 112 128 electron density evidence A Regnase-1 construct consisting of PIN and ZF domains derived from Mus musculus was crystallized; however, the electron density of the ZF domain was low, indicating that the ZF domain is highly mobile in the absence of target mRNA or possibly other protein-protein interactions. DISCUSS | |
| 136 138 ZF structure_element A Regnase-1 construct consisting of PIN and ZF domains derived from Mus musculus was crystallized; however, the electron density of the ZF domain was low, indicating that the ZF domain is highly mobile in the absence of target mRNA or possibly other protein-protein interactions. DISCUSS | |
| 175 177 ZF structure_element A Regnase-1 construct consisting of PIN and ZF domains derived from Mus musculus was crystallized; however, the electron density of the ZF domain was low, indicating that the ZF domain is highly mobile in the absence of target mRNA or possibly other protein-protein interactions. DISCUSS | |
| 188 201 highly mobile protein_state A Regnase-1 construct consisting of PIN and ZF domains derived from Mus musculus was crystallized; however, the electron density of the ZF domain was low, indicating that the ZF domain is highly mobile in the absence of target mRNA or possibly other protein-protein interactions. DISCUSS | |
| 209 219 absence of protein_state A Regnase-1 construct consisting of PIN and ZF domains derived from Mus musculus was crystallized; however, the electron density of the ZF domain was low, indicating that the ZF domain is highly mobile in the absence of target mRNA or possibly other protein-protein interactions. DISCUSS | |
| 227 231 mRNA chemical A Regnase-1 construct consisting of PIN and ZF domains derived from Mus musculus was crystallized; however, the electron density of the ZF domain was low, indicating that the ZF domain is highly mobile in the absence of target mRNA or possibly other protein-protein interactions. DISCUSS | |
| 4 7 NMR experimental_method Our NMR experiments confirmed direct binding of the ZF domain to IL-6 mRNA with a Kd of 10 ± 1.1 μM. Furthermore, an in vitro gel shift assay indicated that Regnase-1 containing the ZF domain enhanced target mRNA-binding, but the protein-RNA complex remained in the bottom of the well without entering into the polyacrylamide gel. DISCUSS | |
| 52 54 ZF structure_element Our NMR experiments confirmed direct binding of the ZF domain to IL-6 mRNA with a Kd of 10 ± 1.1 μM. Furthermore, an in vitro gel shift assay indicated that Regnase-1 containing the ZF domain enhanced target mRNA-binding, but the protein-RNA complex remained in the bottom of the well without entering into the polyacrylamide gel. DISCUSS | |
| 65 69 IL-6 protein_type Our NMR experiments confirmed direct binding of the ZF domain to IL-6 mRNA with a Kd of 10 ± 1.1 μM. Furthermore, an in vitro gel shift assay indicated that Regnase-1 containing the ZF domain enhanced target mRNA-binding, but the protein-RNA complex remained in the bottom of the well without entering into the polyacrylamide gel. DISCUSS | |
| 70 74 mRNA chemical Our NMR experiments confirmed direct binding of the ZF domain to IL-6 mRNA with a Kd of 10 ± 1.1 μM. Furthermore, an in vitro gel shift assay indicated that Regnase-1 containing the ZF domain enhanced target mRNA-binding, but the protein-RNA complex remained in the bottom of the well without entering into the polyacrylamide gel. DISCUSS | |
| 82 84 Kd evidence Our NMR experiments confirmed direct binding of the ZF domain to IL-6 mRNA with a Kd of 10 ± 1.1 μM. Furthermore, an in vitro gel shift assay indicated that Regnase-1 containing the ZF domain enhanced target mRNA-binding, but the protein-RNA complex remained in the bottom of the well without entering into the polyacrylamide gel. DISCUSS | |
| 117 141 in vitro gel shift assay experimental_method Our NMR experiments confirmed direct binding of the ZF domain to IL-6 mRNA with a Kd of 10 ± 1.1 μM. Furthermore, an in vitro gel shift assay indicated that Regnase-1 containing the ZF domain enhanced target mRNA-binding, but the protein-RNA complex remained in the bottom of the well without entering into the polyacrylamide gel. DISCUSS | |
| 157 166 Regnase-1 protein Our NMR experiments confirmed direct binding of the ZF domain to IL-6 mRNA with a Kd of 10 ± 1.1 μM. Furthermore, an in vitro gel shift assay indicated that Regnase-1 containing the ZF domain enhanced target mRNA-binding, but the protein-RNA complex remained in the bottom of the well without entering into the polyacrylamide gel. DISCUSS | |
| 182 184 ZF structure_element Our NMR experiments confirmed direct binding of the ZF domain to IL-6 mRNA with a Kd of 10 ± 1.1 μM. Furthermore, an in vitro gel shift assay indicated that Regnase-1 containing the ZF domain enhanced target mRNA-binding, but the protein-RNA complex remained in the bottom of the well without entering into the polyacrylamide gel. DISCUSS | |
| 208 212 mRNA chemical Our NMR experiments confirmed direct binding of the ZF domain to IL-6 mRNA with a Kd of 10 ± 1.1 μM. Furthermore, an in vitro gel shift assay indicated that Regnase-1 containing the ZF domain enhanced target mRNA-binding, but the protein-RNA complex remained in the bottom of the well without entering into the polyacrylamide gel. DISCUSS | |
| 238 241 RNA chemical Our NMR experiments confirmed direct binding of the ZF domain to IL-6 mRNA with a Kd of 10 ± 1.1 μM. Furthermore, an in vitro gel shift assay indicated that Regnase-1 containing the ZF domain enhanced target mRNA-binding, but the protein-RNA complex remained in the bottom of the well without entering into the polyacrylamide gel. DISCUSS | |
| 28 37 Regnase-1 protein These results indicate that Regnase-1 directly binds to RNA and precipitates under such experimental conditions. DISCUSS | |
| 56 59 RNA chemical These results indicate that Regnase-1 directly binds to RNA and precipitates under such experimental conditions. DISCUSS | |
| 59 78 structural analyses experimental_method Due to this limitation, it is difficult to perform further structural analyses of mRNA-Regnase-1 complexes by X-ray crystallography or NMR. DISCUSS | |
| 82 96 mRNA-Regnase-1 complex_assembly Due to this limitation, it is difficult to perform further structural analyses of mRNA-Regnase-1 complexes by X-ray crystallography or NMR. DISCUSS | |
| 110 131 X-ray crystallography experimental_method Due to this limitation, it is difficult to perform further structural analyses of mRNA-Regnase-1 complexes by X-ray crystallography or NMR. DISCUSS | |
| 135 138 NMR experimental_method Due to this limitation, it is difficult to perform further structural analyses of mRNA-Regnase-1 complexes by X-ray crystallography or NMR. DISCUSS | |
| 24 41 crystal structure evidence The previously reported crystal structure of the Regnase-1 PIN domain derived from Homo sapiens is nearly identical to the one derived from Mus musculus in this study, with a backbone RMSD of 0.2 Å. The amino acid sequences corresponding to PIN (residues 134–295) are the two non-identical residues are substituted with similar amino acids. DISCUSS | |
| 49 58 Regnase-1 protein The previously reported crystal structure of the Regnase-1 PIN domain derived from Homo sapiens is nearly identical to the one derived from Mus musculus in this study, with a backbone RMSD of 0.2 Å. The amino acid sequences corresponding to PIN (residues 134–295) are the two non-identical residues are substituted with similar amino acids. DISCUSS | |
| 59 62 PIN structure_element The previously reported crystal structure of the Regnase-1 PIN domain derived from Homo sapiens is nearly identical to the one derived from Mus musculus in this study, with a backbone RMSD of 0.2 Å. The amino acid sequences corresponding to PIN (residues 134–295) are the two non-identical residues are substituted with similar amino acids. DISCUSS | |
| 83 95 Homo sapiens species The previously reported crystal structure of the Regnase-1 PIN domain derived from Homo sapiens is nearly identical to the one derived from Mus musculus in this study, with a backbone RMSD of 0.2 Å. The amino acid sequences corresponding to PIN (residues 134–295) are the two non-identical residues are substituted with similar amino acids. DISCUSS | |
| 140 152 Mus musculus species The previously reported crystal structure of the Regnase-1 PIN domain derived from Homo sapiens is nearly identical to the one derived from Mus musculus in this study, with a backbone RMSD of 0.2 Å. The amino acid sequences corresponding to PIN (residues 134–295) are the two non-identical residues are substituted with similar amino acids. DISCUSS | |
| 184 188 RMSD evidence The previously reported crystal structure of the Regnase-1 PIN domain derived from Homo sapiens is nearly identical to the one derived from Mus musculus in this study, with a backbone RMSD of 0.2 Å. The amino acid sequences corresponding to PIN (residues 134–295) are the two non-identical residues are substituted with similar amino acids. DISCUSS | |
| 241 244 PIN structure_element The previously reported crystal structure of the Regnase-1 PIN domain derived from Homo sapiens is nearly identical to the one derived from Mus musculus in this study, with a backbone RMSD of 0.2 Å. The amino acid sequences corresponding to PIN (residues 134–295) are the two non-identical residues are substituted with similar amino acids. DISCUSS | |
| 255 262 134–295 residue_range The previously reported crystal structure of the Regnase-1 PIN domain derived from Homo sapiens is nearly identical to the one derived from Mus musculus in this study, with a backbone RMSD of 0.2 Å. The amino acid sequences corresponding to PIN (residues 134–295) are the two non-identical residues are substituted with similar amino acids. DISCUSS | |
| 9 14 mouse taxonomy_domain Both the mouse and human PIN domains form head-to-tail oligomers in three distinct crystal forms. DISCUSS | |
| 19 24 human species Both the mouse and human PIN domains form head-to-tail oligomers in three distinct crystal forms. DISCUSS | |
| 25 28 PIN structure_element Both the mouse and human PIN domains form head-to-tail oligomers in three distinct crystal forms. DISCUSS | |
| 42 54 head-to-tail protein_state Both the mouse and human PIN domains form head-to-tail oligomers in three distinct crystal forms. DISCUSS | |
| 55 64 oligomers oligomeric_state Both the mouse and human PIN domains form head-to-tail oligomers in three distinct crystal forms. DISCUSS | |
| 83 96 crystal forms evidence Both the mouse and human PIN domains form head-to-tail oligomers in three distinct crystal forms. DISCUSS | |
| 42 45 PIN structure_element Rao and co-workers previously argued that PIN dimerization is likely to be a crystallographic artifact with no physiological significance, since monomers were dominant in their analytical ultra-centrifugation experiments. DISCUSS | |
| 145 153 monomers oligomeric_state Rao and co-workers previously argued that PIN dimerization is likely to be a crystallographic artifact with no physiological significance, since monomers were dominant in their analytical ultra-centrifugation experiments. DISCUSS | |
| 177 208 analytical ultra-centrifugation experimental_method Rao and co-workers previously argued that PIN dimerization is likely to be a crystallographic artifact with no physiological significance, since monomers were dominant in their analytical ultra-centrifugation experiments. DISCUSS | |
| 17 31 gel filtration experimental_method In contrast, our gel filtration data, mutational analyses, and NMR spectra all indicate that the PIN domain forms a head-to-tail dimer in solution in a manner similar to the crystal structure. DISCUSS | |
| 38 57 mutational analyses experimental_method In contrast, our gel filtration data, mutational analyses, and NMR spectra all indicate that the PIN domain forms a head-to-tail dimer in solution in a manner similar to the crystal structure. DISCUSS | |
| 63 66 NMR experimental_method In contrast, our gel filtration data, mutational analyses, and NMR spectra all indicate that the PIN domain forms a head-to-tail dimer in solution in a manner similar to the crystal structure. DISCUSS | |
| 67 74 spectra evidence In contrast, our gel filtration data, mutational analyses, and NMR spectra all indicate that the PIN domain forms a head-to-tail dimer in solution in a manner similar to the crystal structure. DISCUSS | |
| 97 100 PIN structure_element In contrast, our gel filtration data, mutational analyses, and NMR spectra all indicate that the PIN domain forms a head-to-tail dimer in solution in a manner similar to the crystal structure. DISCUSS | |
| 116 128 head-to-tail protein_state In contrast, our gel filtration data, mutational analyses, and NMR spectra all indicate that the PIN domain forms a head-to-tail dimer in solution in a manner similar to the crystal structure. DISCUSS | |
| 129 134 dimer oligomeric_state In contrast, our gel filtration data, mutational analyses, and NMR spectra all indicate that the PIN domain forms a head-to-tail dimer in solution in a manner similar to the crystal structure. DISCUSS | |
| 174 191 crystal structure evidence In contrast, our gel filtration data, mutational analyses, and NMR spectra all indicate that the PIN domain forms a head-to-tail dimer in solution in a manner similar to the crystal structure. DISCUSS | |
| 137 145 oligomer oligomeric_state This inconsistency might be due to difference in the analytical methods and/or protein concentrations used in each experiment, since the oligomer formation of PIN was dependent on the protein concentration in our study. DISCUSS | |
| 159 162 PIN structure_element This inconsistency might be due to difference in the analytical methods and/or protein concentrations used in each experiment, since the oligomer formation of PIN was dependent on the protein concentration in our study. DISCUSS | |
| 0 16 Single mutations experimental_method Single mutations to residues involved in the putative oligomeric interaction of PIN monomerized as expected and these mutants lost their RNase activity as well. DISCUSS | |
| 80 83 PIN structure_element Single mutations to residues involved in the putative oligomeric interaction of PIN monomerized as expected and these mutants lost their RNase activity as well. DISCUSS | |
| 84 95 monomerized oligomeric_state Single mutations to residues involved in the putative oligomeric interaction of PIN monomerized as expected and these mutants lost their RNase activity as well. DISCUSS | |
| 118 125 mutants protein_state Single mutations to residues involved in the putative oligomeric interaction of PIN monomerized as expected and these mutants lost their RNase activity as well. DISCUSS | |
| 137 142 RNase protein_type Single mutations to residues involved in the putative oligomeric interaction of PIN monomerized as expected and these mutants lost their RNase activity as well. DISCUSS | |
| 10 13 NMR experimental_method Since the NMR spectra of monomeric mutants overlaps with those of the oligomeric forms, it is unlikely that the tertiary structure of the monomeric mutants were affected by the mutations. (Supplementary Fig. 4b,c). DISCUSS | |
| 14 21 spectra evidence Since the NMR spectra of monomeric mutants overlaps with those of the oligomeric forms, it is unlikely that the tertiary structure of the monomeric mutants were affected by the mutations. (Supplementary Fig. 4b,c). DISCUSS | |
| 25 34 monomeric oligomeric_state Since the NMR spectra of monomeric mutants overlaps with those of the oligomeric forms, it is unlikely that the tertiary structure of the monomeric mutants were affected by the mutations. (Supplementary Fig. 4b,c). DISCUSS | |
| 35 42 mutants protein_state Since the NMR spectra of monomeric mutants overlaps with those of the oligomeric forms, it is unlikely that the tertiary structure of the monomeric mutants were affected by the mutations. (Supplementary Fig. 4b,c). DISCUSS | |
| 138 147 monomeric oligomeric_state Since the NMR spectra of monomeric mutants overlaps with those of the oligomeric forms, it is unlikely that the tertiary structure of the monomeric mutants were affected by the mutations. (Supplementary Fig. 4b,c). DISCUSS | |
| 148 155 mutants protein_state Since the NMR spectra of monomeric mutants overlaps with those of the oligomeric forms, it is unlikely that the tertiary structure of the monomeric mutants were affected by the mutations. (Supplementary Fig. 4b,c). DISCUSS | |
| 47 50 PIN structure_element Based on these observations, we concluded that PIN-PIN dimer formation is critical for Regnase-1 RNase activity in vitro. DISCUSS | |
| 51 54 PIN structure_element Based on these observations, we concluded that PIN-PIN dimer formation is critical for Regnase-1 RNase activity in vitro. DISCUSS | |
| 55 60 dimer oligomeric_state Based on these observations, we concluded that PIN-PIN dimer formation is critical for Regnase-1 RNase activity in vitro. DISCUSS | |
| 87 96 Regnase-1 protein Based on these observations, we concluded that PIN-PIN dimer formation is critical for Regnase-1 RNase activity in vitro. DISCUSS | |
| 97 102 RNase protein_type Based on these observations, we concluded that PIN-PIN dimer formation is critical for Regnase-1 RNase activity in vitro. DISCUSS | |
| 11 28 crystal structure evidence Within the crystal structure of the PIN dimer, the Regnase-1 specific basic regions in both the “primary” and “secondary” PINs are located around the catalytic site of the primary PIN (Supplementary Fig. 6). DISCUSS | |
| 36 39 PIN structure_element Within the crystal structure of the PIN dimer, the Regnase-1 specific basic regions in both the “primary” and “secondary” PINs are located around the catalytic site of the primary PIN (Supplementary Fig. 6). DISCUSS | |
| 40 45 dimer oligomeric_state Within the crystal structure of the PIN dimer, the Regnase-1 specific basic regions in both the “primary” and “secondary” PINs are located around the catalytic site of the primary PIN (Supplementary Fig. 6). DISCUSS | |
| 51 60 Regnase-1 protein Within the crystal structure of the PIN dimer, the Regnase-1 specific basic regions in both the “primary” and “secondary” PINs are located around the catalytic site of the primary PIN (Supplementary Fig. 6). DISCUSS | |
| 97 104 primary protein_state Within the crystal structure of the PIN dimer, the Regnase-1 specific basic regions in both the “primary” and “secondary” PINs are located around the catalytic site of the primary PIN (Supplementary Fig. 6). DISCUSS | |
| 111 120 secondary protein_state Within the crystal structure of the PIN dimer, the Regnase-1 specific basic regions in both the “primary” and “secondary” PINs are located around the catalytic site of the primary PIN (Supplementary Fig. 6). DISCUSS | |
| 122 126 PINs structure_element Within the crystal structure of the PIN dimer, the Regnase-1 specific basic regions in both the “primary” and “secondary” PINs are located around the catalytic site of the primary PIN (Supplementary Fig. 6). DISCUSS | |
| 150 164 catalytic site site Within the crystal structure of the PIN dimer, the Regnase-1 specific basic regions in both the “primary” and “secondary” PINs are located around the catalytic site of the primary PIN (Supplementary Fig. 6). DISCUSS | |
| 180 183 PIN structure_element Within the crystal structure of the PIN dimer, the Regnase-1 specific basic regions in both the “primary” and “secondary” PINs are located around the catalytic site of the primary PIN (Supplementary Fig. 6). DISCUSS | |
| 14 49 structure-based mutational analyses experimental_method Moreover, our structure-based mutational analyses showed these two Regnase-1 specific basic regions were essential for target mRNA cleavage in vitro. DISCUSS | |
| 67 76 Regnase-1 protein Moreover, our structure-based mutational analyses showed these two Regnase-1 specific basic regions were essential for target mRNA cleavage in vitro. DISCUSS | |
| 126 130 mRNA chemical Moreover, our structure-based mutational analyses showed these two Regnase-1 specific basic regions were essential for target mRNA cleavage in vitro. DISCUSS | |
| 4 18 cleavage assay experimental_method The cleavage assay also showed that the NTD is crucial for efficient mRNA cleavage. DISCUSS | |
| 40 43 NTD structure_element The cleavage assay also showed that the NTD is crucial for efficient mRNA cleavage. DISCUSS | |
| 69 73 mRNA chemical The cleavage assay also showed that the NTD is crucial for efficient mRNA cleavage. DISCUSS | |
| 28 31 NTD structure_element Moreover, we found that the NTD associates with the oligomeric surface of the primary PIN, docking to a helix that harbors its catalytic residues (Figs 2b and 3a). DISCUSS | |
| 52 70 oligomeric surface site Moreover, we found that the NTD associates with the oligomeric surface of the primary PIN, docking to a helix that harbors its catalytic residues (Figs 2b and 3a). DISCUSS | |
| 78 85 primary protein_state Moreover, we found that the NTD associates with the oligomeric surface of the primary PIN, docking to a helix that harbors its catalytic residues (Figs 2b and 3a). DISCUSS | |
| 86 89 PIN structure_element Moreover, we found that the NTD associates with the oligomeric surface of the primary PIN, docking to a helix that harbors its catalytic residues (Figs 2b and 3a). DISCUSS | |
| 104 109 helix structure_element Moreover, we found that the NTD associates with the oligomeric surface of the primary PIN, docking to a helix that harbors its catalytic residues (Figs 2b and 3a). DISCUSS | |
| 127 145 catalytic residues site Moreover, we found that the NTD associates with the oligomeric surface of the primary PIN, docking to a helix that harbors its catalytic residues (Figs 2b and 3a). DISCUSS | |
| 39 42 NTD structure_element Taken together, this suggests that the NTD and the PIN domain compete for a common binding site. DISCUSS | |
| 51 54 PIN structure_element Taken together, this suggests that the NTD and the PIN domain compete for a common binding site. DISCUSS | |
| 76 95 common binding site site Taken together, this suggests that the NTD and the PIN domain compete for a common binding site. DISCUSS | |
| 4 12 affinity evidence The affinity of the domain-domain interaction between two PIN domains (Kd = ~10−4 M) is similar to that of the NTD-PIN (Kd = 110 ± 5.8 μM) interactions; however, the covalent connection corresponding to residues 90–133 between the NTD and the primary PIN will greatly enhance the intramolecular domain interaction in the case of full-length Regnase-1. DISCUSS | |
| 58 61 PIN structure_element The affinity of the domain-domain interaction between two PIN domains (Kd = ~10−4 M) is similar to that of the NTD-PIN (Kd = 110 ± 5.8 μM) interactions; however, the covalent connection corresponding to residues 90–133 between the NTD and the primary PIN will greatly enhance the intramolecular domain interaction in the case of full-length Regnase-1. DISCUSS | |
| 71 73 Kd evidence The affinity of the domain-domain interaction between two PIN domains (Kd = ~10−4 M) is similar to that of the NTD-PIN (Kd = 110 ± 5.8 μM) interactions; however, the covalent connection corresponding to residues 90–133 between the NTD and the primary PIN will greatly enhance the intramolecular domain interaction in the case of full-length Regnase-1. DISCUSS | |
| 111 114 NTD structure_element The affinity of the domain-domain interaction between two PIN domains (Kd = ~10−4 M) is similar to that of the NTD-PIN (Kd = 110 ± 5.8 μM) interactions; however, the covalent connection corresponding to residues 90–133 between the NTD and the primary PIN will greatly enhance the intramolecular domain interaction in the case of full-length Regnase-1. DISCUSS | |
| 115 118 PIN structure_element The affinity of the domain-domain interaction between two PIN domains (Kd = ~10−4 M) is similar to that of the NTD-PIN (Kd = 110 ± 5.8 μM) interactions; however, the covalent connection corresponding to residues 90–133 between the NTD and the primary PIN will greatly enhance the intramolecular domain interaction in the case of full-length Regnase-1. DISCUSS | |
| 120 122 Kd evidence The affinity of the domain-domain interaction between two PIN domains (Kd = ~10−4 M) is similar to that of the NTD-PIN (Kd = 110 ± 5.8 μM) interactions; however, the covalent connection corresponding to residues 90–133 between the NTD and the primary PIN will greatly enhance the intramolecular domain interaction in the case of full-length Regnase-1. DISCUSS | |
| 212 218 90–133 residue_range The affinity of the domain-domain interaction between two PIN domains (Kd = ~10−4 M) is similar to that of the NTD-PIN (Kd = 110 ± 5.8 μM) interactions; however, the covalent connection corresponding to residues 90–133 between the NTD and the primary PIN will greatly enhance the intramolecular domain interaction in the case of full-length Regnase-1. DISCUSS | |
| 231 234 NTD structure_element The affinity of the domain-domain interaction between two PIN domains (Kd = ~10−4 M) is similar to that of the NTD-PIN (Kd = 110 ± 5.8 μM) interactions; however, the covalent connection corresponding to residues 90–133 between the NTD and the primary PIN will greatly enhance the intramolecular domain interaction in the case of full-length Regnase-1. DISCUSS | |
| 243 250 primary protein_state The affinity of the domain-domain interaction between two PIN domains (Kd = ~10−4 M) is similar to that of the NTD-PIN (Kd = 110 ± 5.8 μM) interactions; however, the covalent connection corresponding to residues 90–133 between the NTD and the primary PIN will greatly enhance the intramolecular domain interaction in the case of full-length Regnase-1. DISCUSS | |
| 251 254 PIN structure_element The affinity of the domain-domain interaction between two PIN domains (Kd = ~10−4 M) is similar to that of the NTD-PIN (Kd = 110 ± 5.8 μM) interactions; however, the covalent connection corresponding to residues 90–133 between the NTD and the primary PIN will greatly enhance the intramolecular domain interaction in the case of full-length Regnase-1. DISCUSS | |
| 329 340 full-length protein_state The affinity of the domain-domain interaction between two PIN domains (Kd = ~10−4 M) is similar to that of the NTD-PIN (Kd = 110 ± 5.8 μM) interactions; however, the covalent connection corresponding to residues 90–133 between the NTD and the primary PIN will greatly enhance the intramolecular domain interaction in the case of full-length Regnase-1. DISCUSS | |
| 341 350 Regnase-1 protein The affinity of the domain-domain interaction between two PIN domains (Kd = ~10−4 M) is similar to that of the NTD-PIN (Kd = 110 ± 5.8 μM) interactions; however, the covalent connection corresponding to residues 90–133 between the NTD and the primary PIN will greatly enhance the intramolecular domain interaction in the case of full-length Regnase-1. DISCUSS | |
| 74 116 docking and molecular dynamics simulations experimental_method While further analyses are necessary to prove this point, our preliminary docking and molecular dynamics simulations indicate that NTD-binding rearranges the catalytic residues of the PIN domain toward an active conformation suitable for binding Mg2+. DISCUSS | |
| 131 134 NTD structure_element While further analyses are necessary to prove this point, our preliminary docking and molecular dynamics simulations indicate that NTD-binding rearranges the catalytic residues of the PIN domain toward an active conformation suitable for binding Mg2+. DISCUSS | |
| 158 176 catalytic residues site While further analyses are necessary to prove this point, our preliminary docking and molecular dynamics simulations indicate that NTD-binding rearranges the catalytic residues of the PIN domain toward an active conformation suitable for binding Mg2+. DISCUSS | |
| 184 187 PIN structure_element While further analyses are necessary to prove this point, our preliminary docking and molecular dynamics simulations indicate that NTD-binding rearranges the catalytic residues of the PIN domain toward an active conformation suitable for binding Mg2+. DISCUSS | |
| 205 211 active protein_state While further analyses are necessary to prove this point, our preliminary docking and molecular dynamics simulations indicate that NTD-binding rearranges the catalytic residues of the PIN domain toward an active conformation suitable for binding Mg2+. DISCUSS | |
| 246 250 Mg2+ chemical While further analyses are necessary to prove this point, our preliminary docking and molecular dynamics simulations indicate that NTD-binding rearranges the catalytic residues of the PIN domain toward an active conformation suitable for binding Mg2+. DISCUSS | |
| 73 78 Malt1 protein In this context, it is interesting that, in response to TCR stimulation, Malt1 cleaves Regnase-1 at R111 to control immune responses in vivo. DISCUSS | |
| 87 96 Regnase-1 protein In this context, it is interesting that, in response to TCR stimulation, Malt1 cleaves Regnase-1 at R111 to control immune responses in vivo. DISCUSS | |
| 100 104 R111 residue_name_number In this context, it is interesting that, in response to TCR stimulation, Malt1 cleaves Regnase-1 at R111 to control immune responses in vivo. DISCUSS | |
| 52 55 NTD structure_element This result is consistent with a model in which the NTD acts as an enhancer, and cleavage of the linker lowers enzymatic activity dramatically. DISCUSS | |
| 97 103 linker structure_element This result is consistent with a model in which the NTD acts as an enhancer, and cleavage of the linker lowers enzymatic activity dramatically. DISCUSS | |
| 15 49 structural and functional analyses experimental_method Based on these structural and functional analyses of Regnase-1 domain-domain interactions, we performed docking simulations of the NTD, PIN dimer, and IL-6 mRNA. DISCUSS | |
| 53 62 Regnase-1 protein Based on these structural and functional analyses of Regnase-1 domain-domain interactions, we performed docking simulations of the NTD, PIN dimer, and IL-6 mRNA. DISCUSS | |
| 104 123 docking simulations experimental_method Based on these structural and functional analyses of Regnase-1 domain-domain interactions, we performed docking simulations of the NTD, PIN dimer, and IL-6 mRNA. DISCUSS | |
| 131 134 NTD structure_element Based on these structural and functional analyses of Regnase-1 domain-domain interactions, we performed docking simulations of the NTD, PIN dimer, and IL-6 mRNA. DISCUSS | |
| 136 139 PIN structure_element Based on these structural and functional analyses of Regnase-1 domain-domain interactions, we performed docking simulations of the NTD, PIN dimer, and IL-6 mRNA. DISCUSS | |
| 140 145 dimer oligomeric_state Based on these structural and functional analyses of Regnase-1 domain-domain interactions, we performed docking simulations of the NTD, PIN dimer, and IL-6 mRNA. DISCUSS | |
| 151 155 IL-6 protein_type Based on these structural and functional analyses of Regnase-1 domain-domain interactions, we performed docking simulations of the NTD, PIN dimer, and IL-6 mRNA. DISCUSS | |
| 156 160 mRNA chemical Based on these structural and functional analyses of Regnase-1 domain-domain interactions, we performed docking simulations of the NTD, PIN dimer, and IL-6 mRNA. DISCUSS | |
| 37 50 cleavage site site We incorporated information from the cleavage site of IL-6 mRNA in vitro is indicated by denaturing polyacrylamide gel electrophoresis (Supplementary Fig. 7a,b). DISCUSS | |
| 54 58 IL-6 protein_type We incorporated information from the cleavage site of IL-6 mRNA in vitro is indicated by denaturing polyacrylamide gel electrophoresis (Supplementary Fig. 7a,b). DISCUSS | |
| 59 63 mRNA chemical We incorporated information from the cleavage site of IL-6 mRNA in vitro is indicated by denaturing polyacrylamide gel electrophoresis (Supplementary Fig. 7a,b). DISCUSS | |
| 100 134 polyacrylamide gel electrophoresis experimental_method We incorporated information from the cleavage site of IL-6 mRNA in vitro is indicated by denaturing polyacrylamide gel electrophoresis (Supplementary Fig. 7a,b). DISCUSS | |
| 4 11 docking experimental_method The docking result revealed multiple RNA binding modes that satisfied the experimental results in vitro (Supplementary Fig. 7c,d), however, it should be noted that, in vivo, there would likely be many other RNA-binding proteins that would protect loop regions from cleavage by Regnase-1. DISCUSS | |
| 37 40 RNA chemical The docking result revealed multiple RNA binding modes that satisfied the experimental results in vitro (Supplementary Fig. 7c,d), however, it should be noted that, in vivo, there would likely be many other RNA-binding proteins that would protect loop regions from cleavage by Regnase-1. DISCUSS | |
| 207 227 RNA-binding proteins protein_type The docking result revealed multiple RNA binding modes that satisfied the experimental results in vitro (Supplementary Fig. 7c,d), however, it should be noted that, in vivo, there would likely be many other RNA-binding proteins that would protect loop regions from cleavage by Regnase-1. DISCUSS | |
| 247 251 loop structure_element The docking result revealed multiple RNA binding modes that satisfied the experimental results in vitro (Supplementary Fig. 7c,d), however, it should be noted that, in vivo, there would likely be many other RNA-binding proteins that would protect loop regions from cleavage by Regnase-1. DISCUSS | |
| 277 286 Regnase-1 protein The docking result revealed multiple RNA binding modes that satisfied the experimental results in vitro (Supplementary Fig. 7c,d), however, it should be noted that, in vivo, there would likely be many other RNA-binding proteins that would protect loop regions from cleavage by Regnase-1. DISCUSS | |
| 35 44 Regnase-1 protein The overall model of regulation of Regnase-1 RNase activity through domain-domain interactions in vitro is summarized in Fig. 6. DISCUSS | |
| 45 50 RNase protein_type The overall model of regulation of Regnase-1 RNase activity through domain-domain interactions in vitro is summarized in Fig. 6. DISCUSS | |
| 7 17 absence of protein_state In the absence of target mRNA, the PIN domain forms head-to-tail oligomers at high concentration. DISCUSS | |
| 25 29 mRNA chemical In the absence of target mRNA, the PIN domain forms head-to-tail oligomers at high concentration. DISCUSS | |
| 35 38 PIN structure_element In the absence of target mRNA, the PIN domain forms head-to-tail oligomers at high concentration. DISCUSS | |
| 52 64 head-to-tail protein_state In the absence of target mRNA, the PIN domain forms head-to-tail oligomers at high concentration. DISCUSS | |
| 65 74 oligomers oligomeric_state In the absence of target mRNA, the PIN domain forms head-to-tail oligomers at high concentration. DISCUSS | |
| 2 14 fully active protein_state A fully active catalytic center can be formed only when the NTD associates with the oligomer surface of the PIN domain, which terminates the head-to-tail oligomer formation in one direction (primary PIN), and forms a functional dimer together with the neighboring PIN (secondary PIN). DISCUSS | |
| 15 31 catalytic center site A fully active catalytic center can be formed only when the NTD associates with the oligomer surface of the PIN domain, which terminates the head-to-tail oligomer formation in one direction (primary PIN), and forms a functional dimer together with the neighboring PIN (secondary PIN). DISCUSS | |
| 60 63 NTD structure_element A fully active catalytic center can be formed only when the NTD associates with the oligomer surface of the PIN domain, which terminates the head-to-tail oligomer formation in one direction (primary PIN), and forms a functional dimer together with the neighboring PIN (secondary PIN). DISCUSS | |
| 84 92 oligomer oligomeric_state A fully active catalytic center can be formed only when the NTD associates with the oligomer surface of the PIN domain, which terminates the head-to-tail oligomer formation in one direction (primary PIN), and forms a functional dimer together with the neighboring PIN (secondary PIN). DISCUSS | |
| 108 111 PIN structure_element A fully active catalytic center can be formed only when the NTD associates with the oligomer surface of the PIN domain, which terminates the head-to-tail oligomer formation in one direction (primary PIN), and forms a functional dimer together with the neighboring PIN (secondary PIN). DISCUSS | |
| 141 153 head-to-tail protein_state A fully active catalytic center can be formed only when the NTD associates with the oligomer surface of the PIN domain, which terminates the head-to-tail oligomer formation in one direction (primary PIN), and forms a functional dimer together with the neighboring PIN (secondary PIN). DISCUSS | |
| 154 162 oligomer oligomeric_state A fully active catalytic center can be formed only when the NTD associates with the oligomer surface of the PIN domain, which terminates the head-to-tail oligomer formation in one direction (primary PIN), and forms a functional dimer together with the neighboring PIN (secondary PIN). DISCUSS | |
| 191 198 primary protein_state A fully active catalytic center can be formed only when the NTD associates with the oligomer surface of the PIN domain, which terminates the head-to-tail oligomer formation in one direction (primary PIN), and forms a functional dimer together with the neighboring PIN (secondary PIN). DISCUSS | |
| 199 202 PIN structure_element A fully active catalytic center can be formed only when the NTD associates with the oligomer surface of the PIN domain, which terminates the head-to-tail oligomer formation in one direction (primary PIN), and forms a functional dimer together with the neighboring PIN (secondary PIN). DISCUSS | |
| 217 227 functional protein_state A fully active catalytic center can be formed only when the NTD associates with the oligomer surface of the PIN domain, which terminates the head-to-tail oligomer formation in one direction (primary PIN), and forms a functional dimer together with the neighboring PIN (secondary PIN). DISCUSS | |
| 228 233 dimer oligomeric_state A fully active catalytic center can be formed only when the NTD associates with the oligomer surface of the PIN domain, which terminates the head-to-tail oligomer formation in one direction (primary PIN), and forms a functional dimer together with the neighboring PIN (secondary PIN). DISCUSS | |
| 264 267 PIN structure_element A fully active catalytic center can be formed only when the NTD associates with the oligomer surface of the PIN domain, which terminates the head-to-tail oligomer formation in one direction (primary PIN), and forms a functional dimer together with the neighboring PIN (secondary PIN). DISCUSS | |
| 269 278 secondary protein_state A fully active catalytic center can be formed only when the NTD associates with the oligomer surface of the PIN domain, which terminates the head-to-tail oligomer formation in one direction (primary PIN), and forms a functional dimer together with the neighboring PIN (secondary PIN). DISCUSS | |
| 279 282 PIN structure_element A fully active catalytic center can be formed only when the NTD associates with the oligomer surface of the PIN domain, which terminates the head-to-tail oligomer formation in one direction (primary PIN), and forms a functional dimer together with the neighboring PIN (secondary PIN). DISCUSS | |
| 66 75 Regnase-1 protein While further investigations on the domain-domain interactions of Regnase-1 in vivo are necessary, these intramolecular and intermolecular domain interactions of Regnase-1 appear to structurally constrain Regnase-1activity, which, in turn, enables tight regulation of immune responses. DISCUSS | |
| 162 171 Regnase-1 protein While further investigations on the domain-domain interactions of Regnase-1 in vivo are necessary, these intramolecular and intermolecular domain interactions of Regnase-1 appear to structurally constrain Regnase-1activity, which, in turn, enables tight regulation of immune responses. DISCUSS | |
| 205 214 Regnase-1 protein While further investigations on the domain-domain interactions of Regnase-1 in vivo are necessary, these intramolecular and intermolecular domain interactions of Regnase-1 appear to structurally constrain Regnase-1activity, which, in turn, enables tight regulation of immune responses. DISCUSS | |
| 0 34 Structural and functional analyses experimental_method Structural and functional analyses of Regnase-1. FIG | |
| 38 47 Regnase-1 protein Structural and functional analyses of Regnase-1. FIG | |
| 27 36 Regnase-1 protein (a) Domain architecture of Regnase-1. (b) Solution structure of the NTD. (c) Crystal structure of the PIN domain. FIG | |
| 42 60 Solution structure evidence (a) Domain architecture of Regnase-1. (b) Solution structure of the NTD. (c) Crystal structure of the PIN domain. FIG | |
| 68 71 NTD structure_element (a) Domain architecture of Regnase-1. (b) Solution structure of the NTD. (c) Crystal structure of the PIN domain. FIG | |
| 77 94 Crystal structure evidence (a) Domain architecture of Regnase-1. (b) Solution structure of the NTD. (c) Crystal structure of the PIN domain. FIG | |
| 102 105 PIN structure_element (a) Domain architecture of Regnase-1. (b) Solution structure of the NTD. (c) Crystal structure of the PIN domain. FIG | |
| 0 9 Catalytic protein_state Catalytic Asp residues were shown in sticks. FIG | |
| 10 13 Asp residue_name Catalytic Asp residues were shown in sticks. FIG | |
| 4 22 Solution structure evidence (d) Solution structure of the ZF domain. FIG | |
| 30 32 ZF structure_element (d) Solution structure of the ZF domain. FIG | |
| 6 9 Cys residue_name Three Cys residues and one His residue responsible for Zn2+-binding were shown in sticks. FIG | |
| 27 30 His residue_name Three Cys residues and one His residue responsible for Zn2+-binding were shown in sticks. FIG | |
| 4 22 Solution structure evidence (e) Solution structure of the CTD. FIG | |
| 30 33 CTD structure_element (e) Solution structure of the CTD. FIG | |
| 8 18 structures evidence All the structures were colored in rainbow from N-terminus (blue) to C-terminus (red). FIG | |
| 4 36 In vitro gel shift binding assay experimental_method (f) In vitro gel shift binding assay between Regnase-1 and IL-6 mRNA. FIG | |
| 45 54 Regnase-1 protein (f) In vitro gel shift binding assay between Regnase-1 and IL-6 mRNA. FIG | |
| 59 63 IL-6 protein_type (f) In vitro gel shift binding assay between Regnase-1 and IL-6 mRNA. FIG | |
| 64 68 mRNA chemical (f) In vitro gel shift binding assay between Regnase-1 and IL-6 mRNA. FIG | |
| 0 22 Fluorescence intensity evidence Fluorescence intensity of the free IL-6 in each sample was indicated as the percentage against that in the absence of Regnase-1. FIG | |
| 30 34 free protein_state Fluorescence intensity of the free IL-6 in each sample was indicated as the percentage against that in the absence of Regnase-1. FIG | |
| 35 39 IL-6 protein_type Fluorescence intensity of the free IL-6 in each sample was indicated as the percentage against that in the absence of Regnase-1. FIG | |
| 107 117 absence of protein_state Fluorescence intensity of the free IL-6 in each sample was indicated as the percentage against that in the absence of Regnase-1. FIG | |
| 118 127 Regnase-1 protein Fluorescence intensity of the free IL-6 in each sample was indicated as the percentage against that in the absence of Regnase-1. FIG | |
| 15 24 Regnase-1 protein (g) Binding of Regnase-1 and IL-6 mRNA was plotted. FIG | |
| 29 33 IL-6 protein_type (g) Binding of Regnase-1 and IL-6 mRNA was plotted. FIG | |
| 34 38 mRNA chemical (g) Binding of Regnase-1 and IL-6 mRNA was plotted. FIG | |
| 28 32 IL-6 protein_type The percentage of the bound IL-6 was calculated based on the fluorescence intensities of the free IL-6 quantified in (f). FIG | |
| 61 85 fluorescence intensities evidence The percentage of the bound IL-6 was calculated based on the fluorescence intensities of the free IL-6 quantified in (f). FIG | |
| 98 102 IL-6 protein_type The percentage of the bound IL-6 was calculated based on the fluorescence intensities of the free IL-6 quantified in (f). FIG | |
| 4 27 In vitro cleavage assay experimental_method (h) In vitro cleavage assay of Regnase-1 to IL-6 mRNA. FIG | |
| 31 40 Regnase-1 protein (h) In vitro cleavage assay of Regnase-1 to IL-6 mRNA. FIG | |
| 44 48 IL-6 protein_type (h) In vitro cleavage assay of Regnase-1 to IL-6 mRNA. FIG | |
| 49 53 mRNA chemical (h) In vitro cleavage assay of Regnase-1 to IL-6 mRNA. FIG | |
| 0 22 Fluorescence intensity evidence Fluorescence intensity of the uncleaved IL-6 mRNA was indicated as the percentage against that in the absence of Regnase-1. FIG | |
| 30 39 uncleaved protein_state Fluorescence intensity of the uncleaved IL-6 mRNA was indicated as the percentage against that in the absence of Regnase-1. FIG | |
| 40 44 IL-6 protein_type Fluorescence intensity of the uncleaved IL-6 mRNA was indicated as the percentage against that in the absence of Regnase-1. FIG | |
| 45 49 mRNA chemical Fluorescence intensity of the uncleaved IL-6 mRNA was indicated as the percentage against that in the absence of Regnase-1. FIG | |
| 102 112 absence of protein_state Fluorescence intensity of the uncleaved IL-6 mRNA was indicated as the percentage against that in the absence of Regnase-1. FIG | |
| 113 122 Regnase-1 protein Fluorescence intensity of the uncleaved IL-6 mRNA was indicated as the percentage against that in the absence of Regnase-1. FIG | |
| 0 12 Head-to-tail protein_state Head-to-tail oligomer formation of the PIN domain is crucial for the RNase activity of Regnase-1. FIG | |
| 13 21 oligomer oligomeric_state Head-to-tail oligomer formation of the PIN domain is crucial for the RNase activity of Regnase-1. FIG | |
| 39 42 PIN structure_element Head-to-tail oligomer formation of the PIN domain is crucial for the RNase activity of Regnase-1. FIG | |
| 69 74 RNase protein_type Head-to-tail oligomer formation of the PIN domain is crucial for the RNase activity of Regnase-1. FIG | |
| 87 96 Regnase-1 protein Head-to-tail oligomer formation of the PIN domain is crucial for the RNase activity of Regnase-1. FIG | |
| 4 27 Gel filtration analyses experimental_method (a) Gel filtration analyses of the PIN domain. FIG | |
| 35 38 PIN structure_element (a) Gel filtration analyses of the PIN domain. FIG | |
| 4 9 Dimer oligomeric_state (b) Dimer structure of the PIN domain. FIG | |
| 10 19 structure evidence (b) Dimer structure of the PIN domain. FIG | |
| 27 30 PIN structure_element (b) Dimer structure of the PIN domain. FIG | |
| 4 7 PIN structure_element Two PIN molecules in the crystal were colored white and green, respectively. FIG | |
| 25 32 crystal evidence Two PIN molecules in the crystal were colored white and green, respectively. FIG | |
| 0 18 Catalytic residues site Catalytic residues and mutated residues were shown in sticks. FIG | |
| 74 78 R215 residue_name_number Residues important for the oligomeric interaction were colored red, while R215 that was dispensable for the oligomeric interaction was colored blue. (c) RNase activity of monomeric mutants for IL-6 mRNA was analyzed. FIG | |
| 153 158 RNase protein_type Residues important for the oligomeric interaction were colored red, while R215 that was dispensable for the oligomeric interaction was colored blue. (c) RNase activity of monomeric mutants for IL-6 mRNA was analyzed. FIG | |
| 171 180 monomeric oligomeric_state Residues important for the oligomeric interaction were colored red, while R215 that was dispensable for the oligomeric interaction was colored blue. (c) RNase activity of monomeric mutants for IL-6 mRNA was analyzed. FIG | |
| 181 188 mutants protein_state Residues important for the oligomeric interaction were colored red, while R215 that was dispensable for the oligomeric interaction was colored blue. (c) RNase activity of monomeric mutants for IL-6 mRNA was analyzed. FIG | |
| 193 197 IL-6 protein_type Residues important for the oligomeric interaction were colored red, while R215 that was dispensable for the oligomeric interaction was colored blue. (c) RNase activity of monomeric mutants for IL-6 mRNA was analyzed. FIG | |
| 198 202 mRNA chemical Residues important for the oligomeric interaction were colored red, while R215 that was dispensable for the oligomeric interaction was colored blue. (c) RNase activity of monomeric mutants for IL-6 mRNA was analyzed. FIG | |
| 38 41 NTD structure_element Domain-domain interaction between the NTD and the PIN domain. FIG | |
| 50 53 PIN structure_element Domain-domain interaction between the NTD and the PIN domain. FIG | |
| 4 16 NMR analyses experimental_method (a) NMR analyses of the NTD-binding to the PIN domain. FIG | |
| 24 27 NTD structure_element (a) NMR analyses of the NTD-binding to the PIN domain. FIG | |
| 43 46 PIN structure_element (a) NMR analyses of the NTD-binding to the PIN domain. FIG | |
| 73 81 overlaid experimental_method The residues with significant chemical shift changes were labeled in the overlaid spectra (left) and colored red on the surface and ribbon structure of the PIN domain (right). FIG | |
| 82 89 spectra evidence The residues with significant chemical shift changes were labeled in the overlaid spectra (left) and colored red on the surface and ribbon structure of the PIN domain (right). FIG | |
| 156 159 PIN structure_element The residues with significant chemical shift changes were labeled in the overlaid spectra (left) and colored red on the surface and ribbon structure of the PIN domain (right). FIG | |
| 0 3 Pro residue_name Pro and the residues without analysis were colored black and gray, respectively. FIG | |
| 4 16 NMR analyses experimental_method (b) NMR analyses of the PIN-binding to the NTD. FIG | |
| 24 27 PIN structure_element (b) NMR analyses of the PIN-binding to the NTD. FIG | |
| 43 46 NTD structure_element (b) NMR analyses of the PIN-binding to the NTD. FIG | |
| 18 52 significant chemical shift changes evidence The residues with significant chemical shift changes were labeled in the overlaid spectra (left) and colored red, yellow, or green on the surface and ribbon structure of the NTD. FIG | |
| 73 81 overlaid experimental_method The residues with significant chemical shift changes were labeled in the overlaid spectra (left) and colored red, yellow, or green on the surface and ribbon structure of the NTD. FIG | |
| 82 89 spectra evidence The residues with significant chemical shift changes were labeled in the overlaid spectra (left) and colored red, yellow, or green on the surface and ribbon structure of the NTD. FIG | |
| 174 177 NTD structure_element The residues with significant chemical shift changes were labeled in the overlaid spectra (left) and colored red, yellow, or green on the surface and ribbon structure of the NTD. FIG | |
| 0 3 S62 residue_name_number S62 was colored gray and excluded from the analysis, due to low signal intensity. FIG | |
| 25 28 NTD structure_element (c) Docking model of the NTD and the PIN domain. FIG | |
| 37 40 PIN structure_element (c) Docking model of the NTD and the PIN domain. FIG | |
| 4 7 NTD structure_element The NTD and the PIN domain are shown in cyan and white, respectively. FIG | |
| 16 19 PIN structure_element The NTD and the PIN domain are shown in cyan and white, respectively. FIG | |
| 56 73 docking structure evidence Residues in close proximity (<5 Å) to each other in the docking structure were colored yellow. FIG | |
| 0 18 Catalytic residues site Catalytic residues of the PIN domain are shown in sticks, and the residues that exhibited significant chemical shift changes in (a,b) were labeled. FIG | |
| 26 29 PIN structure_element Catalytic residues of the PIN domain are shown in sticks, and the residues that exhibited significant chemical shift changes in (a,b) were labeled. FIG | |
| 90 124 significant chemical shift changes evidence Catalytic residues of the PIN domain are shown in sticks, and the residues that exhibited significant chemical shift changes in (a,b) were labeled. FIG | |
| 25 28 PIN structure_element Critical residues in the PIN domain for the RNase activity of Regnase-1. FIG | |
| 44 49 RNase protein_type Critical residues in the PIN domain for the RNase activity of Regnase-1. FIG | |
| 62 71 Regnase-1 protein Critical residues in the PIN domain for the RNase activity of Regnase-1. FIG | |
| 4 27 In vitro cleavage assay experimental_method (a) In vitro cleavage assay of basic residue mutants for IL-6 mRNA. FIG | |
| 45 52 mutants protein_state (a) In vitro cleavage assay of basic residue mutants for IL-6 mRNA. FIG | |
| 57 61 IL-6 protein_type (a) In vitro cleavage assay of basic residue mutants for IL-6 mRNA. FIG | |
| 62 66 mRNA chemical (a) In vitro cleavage assay of basic residue mutants for IL-6 mRNA. FIG | |
| 4 27 In vitro cleavage assay experimental_method (b) In vitro cleavage assay of basic residue mutants for Regnase-1 mRNA. FIG | |
| 45 52 mutants protein_state (b) In vitro cleavage assay of basic residue mutants for Regnase-1 mRNA. FIG | |
| 57 66 Regnase-1 protein (b) In vitro cleavage assay of basic residue mutants for Regnase-1 mRNA. FIG | |
| 67 71 mRNA chemical (b) In vitro cleavage assay of basic residue mutants for Regnase-1 mRNA. FIG | |
| 4 26 fluorescence intensity evidence The fluorescence intensity of the uncleaved mRNA was quantified and the results were mapped on the PIN dimer structure. FIG | |
| 34 43 uncleaved protein_state The fluorescence intensity of the uncleaved mRNA was quantified and the results were mapped on the PIN dimer structure. FIG | |
| 44 48 mRNA chemical The fluorescence intensity of the uncleaved mRNA was quantified and the results were mapped on the PIN dimer structure. FIG | |
| 99 102 PIN structure_element The fluorescence intensity of the uncleaved mRNA was quantified and the results were mapped on the PIN dimer structure. FIG | |
| 103 108 dimer oligomeric_state The fluorescence intensity of the uncleaved mRNA was quantified and the results were mapped on the PIN dimer structure. FIG | |
| 109 118 structure evidence The fluorescence intensity of the uncleaved mRNA was quantified and the results were mapped on the PIN dimer structure. FIG | |
| 81 86 RNase protein_type Mutated basic residues were shown in sticks and those with significantly reduced RNase activities were colored red or yellow. FIG | |
| 44 53 Regnase-1 protein Heterodimer formation by combination of the Regnase-1 basic residue mutants and the DDNN mutant restored the RNase activity. FIG | |
| 68 75 mutants protein_state Heterodimer formation by combination of the Regnase-1 basic residue mutants and the DDNN mutant restored the RNase activity. FIG | |
| 84 88 DDNN mutant Heterodimer formation by combination of the Regnase-1 basic residue mutants and the DDNN mutant restored the RNase activity. FIG | |
| 89 95 mutant protein_state Heterodimer formation by combination of the Regnase-1 basic residue mutants and the DDNN mutant restored the RNase activity. FIG | |
| 109 114 RNase protein_type Heterodimer formation by combination of the Regnase-1 basic residue mutants and the DDNN mutant restored the RNase activity. FIG | |
| 65 88 In vitro cleavage assay experimental_method (a) Cartoon representation of the concept of the experiment. (b) In vitro cleavage assay of Regnase-1 for IL-6 mRNA. FIG | |
| 92 101 Regnase-1 protein (a) Cartoon representation of the concept of the experiment. (b) In vitro cleavage assay of Regnase-1 for IL-6 mRNA. FIG | |
| 106 110 IL-6 protein_type (a) Cartoon representation of the concept of the experiment. (b) In vitro cleavage assay of Regnase-1 for IL-6 mRNA. FIG | |
| 111 115 mRNA chemical (a) Cartoon representation of the concept of the experiment. (b) In vitro cleavage assay of Regnase-1 for IL-6 mRNA. FIG | |
| 4 27 In vitro cleavage assay experimental_method (c) In vitro cleavage assay of Regnase-1 for Regnase-1 mRNA. FIG | |
| 31 40 Regnase-1 protein (c) In vitro cleavage assay of Regnase-1 for Regnase-1 mRNA. FIG | |
| 45 54 Regnase-1 protein (c) In vitro cleavage assay of Regnase-1 for Regnase-1 mRNA. FIG | |
| 55 59 mRNA chemical (c) In vitro cleavage assay of Regnase-1 for Regnase-1 mRNA. FIG | |
| 4 26 fluorescence intensity evidence The fluorescence intensity of the uncleaved mRNA was quantified and the results were mapped on the PIN dimer. FIG | |
| 34 43 uncleaved protein_state The fluorescence intensity of the uncleaved mRNA was quantified and the results were mapped on the PIN dimer. FIG | |
| 44 48 mRNA chemical The fluorescence intensity of the uncleaved mRNA was quantified and the results were mapped on the PIN dimer. FIG | |
| 99 102 PIN structure_element The fluorescence intensity of the uncleaved mRNA was quantified and the results were mapped on the PIN dimer. FIG | |
| 103 108 dimer oligomeric_state The fluorescence intensity of the uncleaved mRNA was quantified and the results were mapped on the PIN dimer. FIG | |
| 20 25 RNase protein_type The mutations whose RNase activities were not increased in the presence of DDNN mutant were colored in blue on the primary PIN. FIG | |
| 63 74 presence of protein_state The mutations whose RNase activities were not increased in the presence of DDNN mutant were colored in blue on the primary PIN. FIG | |
| 75 79 DDNN mutant The mutations whose RNase activities were not increased in the presence of DDNN mutant were colored in blue on the primary PIN. FIG | |
| 80 86 mutant protein_state The mutations whose RNase activities were not increased in the presence of DDNN mutant were colored in blue on the primary PIN. FIG | |
| 123 126 PIN structure_element The mutations whose RNase activities were not increased in the presence of DDNN mutant were colored in blue on the primary PIN. FIG | |
| 20 25 RNase protein_type The mutations whose RNase activities were restored in the presence of DDNN mutant were colored in red or yellow on the primary PIN. FIG | |
| 58 69 presence of protein_state The mutations whose RNase activities were restored in the presence of DDNN mutant were colored in red or yellow on the primary PIN. FIG | |
| 70 74 DDNN mutant The mutations whose RNase activities were restored in the presence of DDNN mutant were colored in red or yellow on the primary PIN. FIG | |
| 75 81 mutant protein_state The mutations whose RNase activities were restored in the presence of DDNN mutant were colored in red or yellow on the primary PIN. FIG | |
| 127 130 PIN structure_element The mutations whose RNase activities were restored in the presence of DDNN mutant were colored in red or yellow on the primary PIN. FIG | |
| 46 55 Regnase-1 protein Schematic representation of regulation of the Regnase-1 catalytic activity through the domain-domain interactions. FIG | |