title: PUF60: a novel U2AF65-related splicing activity.
abstract: We have identified a new pyrimidine-tract binding factor, PUF, that is required, together with U2AF, for efficient reconstitution of RNA splicing in vitro. The activity has been purified and consists of two proteins, PUF60 and the previously described splicing factor p54. p54 and PUF60 form a stable complex in vitro when cotranslated in a reaction mixture. PUF activity, in conjunction with U2AF, facilitates the association of U2 snRNP with the pre-mRNA. This reaction is dependent upon the presence of the large subunit of U2AF, U2AF65, but not the small subunit U2AF35. PUF60 is homologous to both U2AF65 and the yeast splicing factor Mud2p. The C-terminal domain of PUF60, the PUMP domain, is distantly related to the RNA-recognition motif domain, and is probably important in protein-protein interactions.
authors.affiliation: Center for Cancer Research and Department of Biology, Massachusetts Institute of Technology, Cambridge 02139, USA.
authors.forename: P S
authors.initials: PS
authors.lastname: Page-McCaw
authors.forename: K
authors.initials: K
authors.lastname: Amonlirdviman
authors.forename: P A
authors.initials: PA
authors.lastname: Sharp
mesh_terms: D000595:Amino Acid Sequence; D002467:Cell Nucleus / Q000378:metabolism; D002846:Chromatography, Affinity; D006367:HeLa Cells; D006801:Humans; D008969:Molecular Sequence Data; D020559:Monomeric GTP-Binding Proteins; D054778:NM23 Nucleoside Diphosphate Kinases; D009687:Nuclear Proteins / Q000378:metabolism; D009701:Nucleoside-Diphosphate Kinase; D011072:Poly U; D012326:RNA Splicing; D016601:RNA-Binding Proteins / Q000737:chemistry / Q000378:metabolism*; D012261:Ribonucleoproteins / Q000378:metabolism*; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid; D000068103:Serine-Arginine Splicing Factors; D000072668:Splicing Factor U2AF; D014157:Transcription Factors / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*
chemical_list: D054778:NM23 Nucleoside Diphosphate Kinases; D009687:Nuclear Proteins; D016601:RNA-Binding Proteins; D012261:Ribonucleoproteins; C101745:Sfrs11 protein; D000072668:Splicing Factor U2AF; D014157:Transcription Factors; C000607494:U2AF2 protein, human; D000068103:Serine-Arginine Splicing Factors; D011072:Poly U; D009701:Nucleoside-Diphosphate Kinase; D020559:Monomeric GTP-Binding Proteins