title: Splicing inhibition of U2AF65 leads to alternative exon skipping.
abstract: U2 snRNP auxiliary factor 65 kDa (U2AF(65)) is a general splicing factor that contacts polypyrimidine (Py) tract and promotes prespliceosome assembly. In this report, we show that U2AF(65) stimulates alternative exon skipping in spinal muscular atrophy (SMA)-related survival motor neuron (SMN) pre-mRNA. A stronger 5' splice-site mutation of alternative exon abolishes the stimulatory effects of U2AF(65). U2AF(65) overexpression promotes its own binding only on the weaker, not the stronger, Py tract. We further demonstrate that U2AF(65) inhibits splicing of flanking introns of alternative exon in both three-exon and two-exon contexts. Similar U2AF(65) effects were observed in Fas (Apo-1/CD95) pre-mRNA. Strikingly, we demonstrate that U2AF(65) even inhibits general splicing of adenovirus major late (Ad ML) or β-globin pre-mRNA. Thus, we conclude that U2AF(65) possesses a splicing Inhibitory function that leads to alternative exon skipping.
authors.affiliation: School of Life Sciences, Gwangju Institute of Science and Technology, Gwangju 500-712, Korea;
authors.forename: Sunghee
authors.initials: S
authors.lastname: Cho
authors.affiliation: School of Life Sciences, Gwangju Institute of Science and Technology, Gwangju 500-712, Korea;
authors.forename: Heegyum
authors.initials: H
authors.lastname: Moon
authors.affiliation: School of Life Sciences, Gwangju Institute of Science and Technology, Gwangju 500-712, Korea;
authors.forename: Tiing Jen
authors.initials: TJ
authors.lastname: Loh
authors.affiliation: School of Life Sciences, Gwangju Institute of Science and Technology, Gwangju 500-712, Korea;
authors.forename: Ha Na
authors.initials: HN
authors.lastname: Jang
authors.affiliation: School of Life Sciences, Gwangju Institute of Science and Technology, Gwangju 500-712, Korea;
authors.forename: Yongchao
authors.initials: Y
authors.lastname: Liu
authors.affiliation: JiangSu Key Laboratory of Neuroregeneration, Nantong University, Nantong 226001, China;
authors.forename: Jianhua
authors.initials: J
authors.lastname: Zhou
authors.affiliation: Department of Cellular & Molecular Medicine, College of Medicine, Chosun University, Gwangju 501-759, Korea.
authors.forename: Takbum
authors.initials: T
authors.lastname: Ohn
authors.affiliation: School of Life Sciences, Gwangju Institute of Science and Technology, Gwangju 500-712, Korea;
authors.forename: Xuexiu
authors.initials: X
authors.lastname: Zheng
authors.affiliation: School of Life Sciences, Gwangju Institute of Science and Technology, Gwangju 500-712, Korea; haihongshen@gist.ac.kr.
authors.forename: Haihong
authors.initials: H
authors.lastname: Shen
mesh_terms: D017398:Alternative Splicing / Q000235:genetics*; D001483:Base Sequence; D004268:DNA-Binding Proteins / Q000235:genetics; D005091:Exons / Q000235:genetics*; D057809:HEK293 Cells; D006801:Humans; D007438:Introns / Q000235:genetics; D008969:Molecular Sequence Data; D009687:Nuclear Proteins / Q000737:chemistry / Q000235:genetics*; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D012322:RNA Precursors / Q000235:genetics / Q000378:metabolism; D022821:RNA Splice Sites / Q000235:genetics; D012261:Ribonucleoproteins / Q000737:chemistry / Q000235:genetics*; D055532:SMN Complex Proteins / Q000235:genetics; D000072668:Splicing Factor U2AF; D013329:Structure-Activity Relationship; D014157:Transcription Factors / Q000235:genetics; D014764:Viral Proteins / Q000235:genetics; D055544:beta-Globins / Q000235:genetics; D016875:tau Proteins / Q000235:genetics
chemical_list: D004268:DNA-Binding Proteins; C051096:MLTF protein, adenovirus; D009687:Nuclear Proteins; D012322:RNA Precursors; D022821:RNA Splice Sites; D012261:Ribonucleoproteins; D055532:SMN Complex Proteins; D000072668:Splicing Factor U2AF; D014157:Transcription Factors; C000607494:U2AF2 protein, human; D014764:Viral Proteins; D055544:beta-Globins; D016875:tau Proteins
keywords: SMN; U2AF65; exon exclusion; pre-mRNA splicing; splicing inhibition