TY  - JOUR
A1  - Kuwasako, Kanako
A1  - Namek, Nobukazu
A1  - Tsuda, Kengo
A1  - Takahashi, Mari
A1  - Sato, Atsuko
A1  - Tochio, Naoya
A1  - Inoue, Makoto
A1  - Terada, Takaho
A1  - Kigawa, Takanori
A1  - Kobayashi, Naohiro
A1  - Shirouzu, Mikako
A1  - Ito, Takuhiro
A1  - Sakamoto, Taiichi
A1  - Wakamatsu, Kaori
A1  - Güntert, Peter
A1  - Takahashi, Seizo
A1  - Yokoyama, Shigeyuki
A1  - Muto, Yutaka
T1  - Solution structure of the first RNA recognition motif domain of human spliceosomal protein SF3b49 and its mode of interaction with a SF3b145 fragment
T2  - Protein science
N2  - The spliceosomal protein SF3b49, a component of the splicing factor 3b (SF3b) protein complex in the U2 small nuclear ribonucleoprotein, contains two RNA recognition motif (RRM) domains. In yeast, the first RRM domain (RRM1) of Hsh49 protein (yeast orthologue of human SF3b49) reportedly interacts with another component, Cus1 protein (orthologue of human SF3b145). Here, we solved the solution structure of the RRM1 of human SF3b49 and examined its mode of interaction with a fragment of human SF3b145 using NMR methods. Chemical shift mapping showed that the SF3b145 fragment spanning residues 598-631 interacts with SF3b49 RRM1, which adopts a canonical RRM fold with a topology of β1-α1-β2-β3-α2-β4. Furthermore, a docking model based on NOESY measurements suggests that residues 607-616 of the SF3b145 fragment adopt a helical structure that binds to RRM1 predominantly via α1, consequently exhibiting a helix-helix interaction in almost antiparallel. This mode of interaction was confirmed by a mutational analysis using GST pull-down assays. Comparison with structures of all RRM domains when complexed with a peptide found that this helix-helix interaction is unique to SF3b49 RRM1. Additionally, all amino acid residues involved in the interaction are well conserved among eukaryotes, suggesting evolutionary conservation of this interaction mode between SF3b49 RRM1 and SF3b145.
KW  - RNA recognition motif
KW  - SF3b145
KW  - SF3b49
KW  - U2 snRNP
KW  - nuclear magnetic resonance
Y1  - 2017
UR  - http://publikationen.ub.uni-frankfurt.de/frontdoor/index/index/docId/43838
UR  - https://nbn-resolving.org/urn:nbn:de:hebis:30:3-438382
SN  - 1469-896X
SN  - 0961-8368
N1  - This is an open access article under the terms of the Creative Commons Attribution-NonCommercial-NoDerivs License, which permits use and distribution in any medium, provided the original work is properly cited, the use is non-commercial and no modifications or adaptations are made.
VL  - 26
IS  - 2
SP  - 280
EP  - 291
PB  - Wiley ; Protein Society
CY  - Hoboken, NJ :  Bethesda, Md.
ER  -