Kanako Kuwasako, Nobukazu Namek, Kengo Tsuda, Mari Takahashi, Atsuko Sato, Naoya Tochio, Makoto Inoue, Takaho Terada, Takanori Kigawa, Naohiro Kobayashi, Mikako Shirouzu, Takuhiro Ito, Taiichi Sakamoto, Kaori Wakamatsu, Peter Güntert, Seizo Takahashi, Shigeyuki Yokoyama, Yutaka Muto
- 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.
MetadatenAuthor: | Kanako KuwasakoORCiD, Nobukazu Namek, Kengo Tsuda, Mari Takahashi, Atsuko Sato, Naoya Tochio, Makoto Inoue, Takaho Terada, Takanori Kigawa, Naohiro Kobayashi, Mikako Shirouzu, Takuhiro Ito, Taiichi Sakamoto, Kaori Wakamatsu, Peter GüntertORCiDGND, Seizo Takahashi, Shigeyuki Yokoyama, Yutaka Muto |
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URN: | urn:nbn:de:hebis:30:3-438382 |
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DOI: | https://doi.org/10.1002/pro.3080 |
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ISSN: | 1469-896X |
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ISSN: | 0961-8368 |
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Pubmed Id: | https://pubmed.ncbi.nlm.nih.gov/27862552 |
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Parent Title (English): | Protein science |
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Publisher: | Wiley ; Protein Society |
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Place of publication: | Hoboken, NJ : Bethesda, Md. |
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Document Type: | Article |
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Language: | English |
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Date of Publication (online): | 2017/09/04 |
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Year of first Publication: | 2016 |
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Publishing Institution: | Universitätsbibliothek Johann Christian Senckenberg |
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Release Date: | 2017/09/04 |
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Tag: | RNA recognition motif; SF3b145; SF3b49; U2 snRNP; nuclear magnetic resonance |
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Volume: | 26 |
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Issue: | 2 |
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Page Number: | 12 |
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First Page: | 280 |
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Last Page: | 291 |
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Note: | 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. |
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HeBIS-PPN: | 428739458 |
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Institutes: | Biochemie, Chemie und Pharmazie / Biochemie und Chemie |
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| Wissenschaftliche Zentren und koordinierte Programme / Zentrum für Biomolekulare Magnetische Resonanz (BMRZ) |
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Dewey Decimal Classification: | 5 Naturwissenschaften und Mathematik / 57 Biowissenschaften; Biologie / 570 Biowissenschaften; Biologie |
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Sammlungen: | Universitätspublikationen |
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Licence (German): | Creative Commons - Namensnennung-Nicht kommerziell - Keine Bearbeitung 4.0 |
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