title: Structural basis for the regulation of enzymatic activity of Regnase-1 by domain-domain interactions.
abstract: 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. 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). The PIN domain harbors the RNase catalytic center; however, it is insufficient for enzymatic activity. We found that the NTD associates with the PIN domain and significantly enhances its RNase activity. The PIN domain forms a head-to-tail oligomer and the dimer interface overlaps with the NTD binding site. Interestingly, mutations blocking PIN oligomerization had no RNase activity, indicating that both oligomerization and NTD binding are crucial for RNase activity in vitro. These results suggest that Regnase-1 RNase activity is tightly controlled by both intramolecular (NTD-PIN) and intermolecular (PIN-PIN) interactions.
authors.affiliation: Faculty of Advanced Life Science, Hokkaido University, Sapporo 001-0021, Japan.
authors.forename: Mariko
authors.initials: M
authors.lastname: Yokogawa
authors.affiliation: Graduate school of Life Science, Hokkaido University, Sapporo 060-0810, Japan.
authors.forename: Takashi
authors.initials: T
authors.lastname: Tsushima
authors.affiliation: Institute of Microbial Chemistry, Microbial Chemistry Research Foundation, Tokyo 141-0021, Japan.
authors.forename: Nobuo N
authors.initials: NN
authors.lastname: Noda
authors.affiliation: Faculty of Advanced Life Science, Hokkaido University, Sapporo 001-0021, Japan.
authors.forename: Hiroyuki
authors.initials: H
authors.lastname: Kumeta
authors.affiliation: Faculty of Advanced Life Science, Hokkaido University, Sapporo 001-0021, Japan.
authors.forename: Yoshiaki
authors.initials: Y
authors.lastname: Enokizono
authors.affiliation: World Premier International Immunology Frontier Research Center, Osaka University, Osaka 565-0871, Japan.
authors.forename: Kazuo
authors.initials: K
authors.lastname: Yamashita
authors.affiliation: World Premier International Immunology Frontier Research Center, Osaka University, Osaka 565-0871, Japan.|Institute for Virus Research, Kyoto University, Kyoto 606-8507, Japan.
authors.forename: Daron M
authors.initials: DM
authors.lastname: Standley
authors.affiliation: World Premier International Immunology Frontier Research Center, Osaka University, Osaka 565-0871, Japan.|Research Institute for Microbial Diseases, Osaka University, Osaka 565-0871, Japan.|Institute for Virus Research, Kyoto University, Kyoto 606-8507, Japan.
authors.forename: Osamu
authors.initials: O
authors.lastname: Takeuchi
authors.affiliation: World Premier International Immunology Frontier Research Center, Osaka University, Osaka 565-0871, Japan.|Research Institute for Microbial Diseases, Osaka University, Osaka 565-0871, Japan.
authors.forename: Shizuo
authors.initials: S
authors.lastname: Akira
authors.affiliation: Faculty of Advanced Life Science, Hokkaido University, Sapporo 001-0021, Japan.
authors.forename: Fuyuhiko
authors.initials: F
authors.lastname: Inagaki
mesh_terms: D000818:Animals; D001665:Binding Sites / Q000235:genetics; D018360:Crystallography, X-Ray; D007249:Inflammation / Q000235:genetics*; D051379:Mice; D008958:Models, Molecular; D009154:Mutation / Q000235:genetics; D011485:Protein Binding; D011487:Protein Conformation; D000072417:Protein Domains / Q000235:genetics; D015202:Protein Engineering; D055503:Protein Multimerization / Q000235:genetics; D012260:Ribonucleases / Q000235:genetics / Q000378:metabolism*; D013329:Structure-Activity Relationship
chemical_list: D012260:Ribonucleases; C541320:Zc3h12a protein, mouse