title: RBFOX and SUP-12 sandwich a G base to cooperatively regulate tissue-specific splicing.
abstract: Tissue-specific alternative pre-mRNA splicing is often cooperatively regulated by multiple splicing factors, but the structural basis of cooperative RNA recognition is poorly understood. In Caenorhabditis elegans, ligand binding specificity of fibroblast growth factor receptors (FGFRs) is determined by mutually exclusive alternative splicing of the sole FGFR gene, egl-15. Here we determined the solution structure of a ternary complex of the RNA-recognition motif (RRM) domains from the RBFOX protein ASD-1, SUP-12 and their target RNA from egl-15. The two RRM domains cooperatively interact with the RNA by sandwiching a G base to form the stable complex. Multichromatic fluorescence splicing reporters confirmed the requirement of the G and the juxtaposition of the respective cis elements for effective splicing regulation in vivo. Moreover, we identified a new target for the heterologous complex through an element search, confirming the functional significance of the intermolecular coordination.
authors.affiliation: 1] RIKEN Systems and Structural Biology Center, Yokohama, Japan. [2] RIKEN Center for Life Science Technologies, Yokohama, Japan. [3] Faculty of Pharmacy and Research Institute of Pharmaceutical Sciences, Musashino University, Nishitokyo, Japan. [4].
authors.forename: Kanako
authors.initials: K
authors.lastname: Kuwasako
authors.affiliation: 1] RIKEN Systems and Structural Biology Center, Yokohama, Japan. [2] RIKEN Center for Life Science Technologies, Yokohama, Japan. [3].
authors.forename: Mari
authors.initials: M
authors.lastname: Takahashi
authors.affiliation: Graduate School of Nanobioscience, Yokohama City University, Yokohama, Japan.
authors.forename: Satoru
authors.initials: S
authors.lastname: Unzai
authors.affiliation: 1] RIKEN Systems and Structural Biology Center, Yokohama, Japan. [2] RIKEN Center for Life Science Technologies, Yokohama, Japan.
authors.forename: Kengo
authors.initials: K
authors.lastname: Tsuda
authors.affiliation: RIKEN Systems and Structural Biology Center, Yokohama, Japan.
authors.forename: Seiko
authors.initials: S
authors.lastname: Yoshikawa
authors.affiliation: RIKEN Systems and Structural Biology Center, Yokohama, Japan.
authors.forename: Fahu
authors.initials: F
authors.lastname: He
authors.affiliation: 1] RIKEN Systems and Structural Biology Center, Yokohama, Japan. [2] Institute for Protein Research, Osaka University, Suita, Japan.
authors.forename: Naohiro
authors.initials: N
authors.lastname: Kobayashi
authors.affiliation: 1] Tatsuo Miyazawa Memorial Program, RIKEN Genomic Sciences Center, Yokohama, Japan. [2] Institute of Biophysical Chemistry, Center for Biomolecular Magnetic Resonance, Goethe University Frankfurt, Frankfurt am Main, Germany. [3] Frankfurt Institute of Advanced Studies, Goethe University Frankfurt, Frankfurt am Main, Germany. [4] Department of Chemistry, Graduate School of Science and Engineering, Tokyo Metropolitan University, Hachioji, Japan.
authors.forename: Peter
authors.initials: P
authors.lastname: Güntert
authors.affiliation: 1] RIKEN Systems and Structural Biology Center, Yokohama, Japan. [2] RIKEN Center for Life Science Technologies, Yokohama, Japan.
authors.forename: Mikako
authors.initials: M
authors.lastname: Shirouzu
authors.affiliation: 1] RIKEN Systems and Structural Biology Center, Yokohama, Japan. [2] RIKEN Center for Life Science Technologies, Yokohama, Japan.
authors.forename: Takuhiro
authors.initials: T
authors.lastname: Ito
authors.affiliation: 1] RIKEN Systems and Structural Biology Center, Yokohama, Japan. [2] RIKEN Center for Life Science Technologies, Yokohama, Japan.
authors.forename: Akiko
authors.initials: A
authors.lastname: Tanaka
authors.affiliation: 1] RIKEN Systems and Structural Biology Center, Yokohama, Japan. [2] RIKEN Structural Biology Laboratory, Yokohama, Japan.
authors.forename: Shigeyuki
authors.initials: S
authors.lastname: Yokoyama
authors.affiliation: 1] Graduate School of Biomedical Science, Tokyo Medical and Dental University, Tokyo, Japan. [2] Department of Anatomy and Developmental Biology, Graduate School of Medicine, Kyoto University, Kyoto, Japan.
authors.forename: Masatoshi
authors.initials: M
authors.lastname: Hagiwara
authors.affiliation: 1] Graduate School of Biomedical Science, Tokyo Medical and Dental University, Tokyo, Japan. [2] Medical Research Institute, Tokyo Medical and Dental University, Tokyo, Japan.
authors.forename: Hidehito
authors.initials: H
authors.lastname: Kuroyanagi
authors.affiliation: 1] RIKEN Systems and Structural Biology Center, Yokohama, Japan. [2] RIKEN Center for Life Science Technologies, Yokohama, Japan. [3] Faculty of Pharmacy and Research Institute of Pharmaceutical Sciences, Musashino University, Nishitokyo, Japan.
authors.forename: Yutaka
authors.initials: Y
authors.lastname: Muto
mesh_terms: D017398:Alternative Splicing; D000818:Animals; D001483:Base Sequence; D017173:Caenorhabditis elegans / Q000737:chemistry / Q000378:metabolism*; D029742:Caenorhabditis elegans Proteins / Q000737:chemistry / Q000378:metabolism*; D018360:Crystallography, X-Ray; D008958:Models, Molecular; D017434:Protein Structure, Tertiary; D012313:RNA / Q000737:chemistry / Q000378:metabolism*; D016601:RNA-Binding Proteins / Q000737:chemistry / Q000378:metabolism*
chemical_list: C515945:Asd-1 protein, C elegans; D029742:Caenorhabditis elegans Proteins; D016601:RNA-Binding Proteins; C525043:Sup-12 protein, C elegans; D012313:RNA