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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 82106 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 82106 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 82106 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 82106 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 82106 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 82106 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 82106 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 82106 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 82106 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 82106 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 82106 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 82106 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 82106 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 82106 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 82106 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 82106 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 82106 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 82106 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 82106 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 82106 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 20200 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 20200 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 20200 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 20200 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 134295) 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 134295) 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 134295) 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 134295) 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 134295) 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 134295) 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 134295) are the two non-identical residues are substituted with similar amino acids. DISCUSS
255 262 134295 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 134295) 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 = ~104 M) is similar to that of the NTD-PIN (Kd = 110 ± 5.8 μM) interactions; however, the covalent connection corresponding to residues 90133 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 = ~104 M) is similar to that of the NTD-PIN (Kd = 110 ± 5.8 μM) interactions; however, the covalent connection corresponding to residues 90133 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 = ~104 M) is similar to that of the NTD-PIN (Kd = 110 ± 5.8 μM) interactions; however, the covalent connection corresponding to residues 90133 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 = ~104 M) is similar to that of the NTD-PIN (Kd = 110 ± 5.8 μM) interactions; however, the covalent connection corresponding to residues 90133 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 = ~104 M) is similar to that of the NTD-PIN (Kd = 110 ± 5.8 μM) interactions; however, the covalent connection corresponding to residues 90133 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 = ~104 M) is similar to that of the NTD-PIN (Kd = 110 ± 5.8 μM) interactions; however, the covalent connection corresponding to residues 90133 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 90133 residue_range The affinity of the domain-domain interaction between two PIN domains (Kd = ~104 M) is similar to that of the NTD-PIN (Kd = 110 ± 5.8 μM) interactions; however, the covalent connection corresponding to residues 90133 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 = ~104 M) is similar to that of the NTD-PIN (Kd = 110 ± 5.8 μM) interactions; however, the covalent connection corresponding to residues 90133 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 = ~104 M) is similar to that of the NTD-PIN (Kd = 110 ± 5.8 μM) interactions; however, the covalent connection corresponding to residues 90133 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 = ~104 M) is similar to that of the NTD-PIN (Kd = 110 ± 5.8 μM) interactions; however, the covalent connection corresponding to residues 90133 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 = ~104 M) is similar to that of the NTD-PIN (Kd = 110 ± 5.8 μM) interactions; however, the covalent connection corresponding to residues 90133 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 = ~104 M) is similar to that of the NTD-PIN (Kd = 110 ± 5.8 μM) interactions; however, the covalent connection corresponding to residues 90133 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