title: Induced folding of the U2AF35 RRM upon binding to U2AF65.
abstract: The human essential splicing factor U2AF (U2 auxiliary factor) consists of 35 and 65 kDa subunits which form a highly stable heterodimer in solution. Copurification of the recombinant U2AF35 RNA recognition motif (U2AF35 RRM) and full-length U2AF65 yields a soluble and functionally active minimal U2AF heterodimer. Recombinant U2AF35 RRM protein free and in complex with three different regions of U2AF65 was characterized by nuclear magnetic resonance spectroscopy. We found that the recombinant U2AF35 RRM is unstructured in solution but its tertiary structure is induced upon binding to U2AF65. This interaction is mediated by the N-terminal proline-rich region of U2AF65 and does not involve the U2AF65 RRMs.
authors.affiliation: Structural and Computational Biology Programme, European Molecular Biology Laboratory, Meyerhofstrasse 1, 69117 Heidelberg, Germany. ekellen@aspirine.u-strasbg.fr
authors.forename: Esther
authors.initials: E
authors.lastname: Kellenberger
authors.forename: Gunter
authors.initials: G
authors.lastname: Stier
authors.forename: Michael
authors.initials: M
authors.lastname: Sattler
mesh_terms: D020816:Amino Acid Motifs; D000595:Amino Acid Sequence; D019281:Dimerization; D006801:Humans; D066298:In Vitro Techniques; D008958:Models, Molecular; D008969:Molecular Sequence Data; D019906:Nuclear Magnetic Resonance, Biomolecular; D009687:Nuclear Proteins; D011485:Protein Binding; D017510:Protein Folding; D017434:Protein Structure, Tertiary; D021122:Protein Subunits; D012326:RNA Splicing; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D012261:Ribonucleoproteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D017386:Sequence Homology, Amino Acid; D000072668:Splicing Factor U2AF
chemical_list: D009687:Nuclear Proteins; D021122:Protein Subunits; D011994:Recombinant Proteins; D012261:Ribonucleoproteins; D000072668:Splicing Factor U2AF; C076753:U2AF1 protein, human; C000607494:U2AF2 protein, human