title: Interaction of U2AF65 RS region with pre-mRNA branch point and promotion of base pairing with U2 snRNA [corrected].
abstract: The mammalian splicing factor U2AF65 binds to the polypyrimidine tract adjacent to the 3' splice site and promotes assembly of U2 small nuclear ribonucleoprotein on the upstream branch point, an interaction that involves base pairing with U2 small nuclear RNA (snRNA). U2AF65 contains an RNA binding domain, required for interaction with the polypyrimidine tract, and an arginine-serine-rich (RS) region, required for U2 snRNP recruitment and splicing. Here it is reported that binding of U2AF65 to the polypyrimidine tract directed the RS domain to contact the branch point and promoted U2 snRNA-branch point base pairing even in the absence of other splicing factors. Analysis of RS domain mutants indicated that the ability of U2AF65 to contact the branch point, to promote the U2 snRNA-branch point interaction, and to support splicing are related activities, requiring only a few basic amino acids. Thus, the U2AF65 RS domain plays a direct role in modulating spliceosomal RNA-RNA interactions.
authors.affiliation: Howard Hughes Medical Institute, University of Massachusetts Medical Center, Worcester, MA 01605, USA.
authors.forename: J
authors.initials: J
authors.lastname: Valcárcel
authors.forename: R K
authors.initials: RK
authors.lastname: Gaur
authors.forename: R
authors.initials: R
authors.lastname: Singh
authors.forename: M R
authors.initials: MR
authors.lastname: Green
mesh_terms: D000595:Amino Acid Sequence; D001120:Arginine / Q000737:chemistry; D001482:Base Composition; D001483:Base Sequence; D003432:Cross-Linking Reagents; D008957:Models, Genetic; D008969:Molecular Sequence Data; D009687:Nuclear Proteins; D009690:Nucleic Acid Conformation; D012322:RNA Precursors / Q000378:metabolism*; D012326:RNA Splicing; D012333:RNA, Messenger / Q000378:metabolism*; D012342:RNA, Small Nuclear / Q000378:metabolism*; D012261:Ribonucleoproteins / Q000737:chemistry / Q000378:metabolism*; D012694:Serine / Q000737:chemistry; D017381:Spliceosomes / Q000378:metabolism; D000072668:Splicing Factor U2AF
chemical_list: D003432:Cross-Linking Reagents; D009687:Nuclear Proteins; D012322:RNA Precursors; D012333:RNA, Messenger; D012342:RNA, Small Nuclear; D012261:Ribonucleoproteins; D000072668:Splicing Factor U2AF; D012694:Serine; D001120:Arginine