annotation_CSV/PMC4772114.csv

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