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