title: RNA-Seq analysis in mutant zebrafish reveals role of U1C protein in alternative splicing regulation.
abstract: Precise 5' splice-site recognition is essential for both constitutive and regulated pre-mRNA splicing. The U1 small nuclear ribonucleoprotein particle (snRNP)-specific protein U1C is involved in this first step of spliceosome assembly and important for stabilizing early splicing complexes. We used an embryonically lethal U1C mutant zebrafish, hi1371, to investigate the potential genomewide role of U1C for splicing regulation. U1C mutant embryos contain overall stable, but U1C-deficient U1 snRNPs. Surprisingly, genomewide RNA-Seq analysis of mutant versus wild-type embryos revealed a large set of specific target genes that changed their alternative splicing patterns in the absence of U1C. Injection of ZfU1C cRNA into mutant embryos and in vivo splicing experiments in HeLa cells after siRNA-mediated U1C knockdown confirmed the U1C dependency and specificity, as well as the functional conservation of the effects observed. In addition, sequence motif analysis of the U1C-dependent 5' splice sites uncovered an association with downstream intronic U-rich elements. In sum, our findings provide evidence for a new role of a general snRNP protein, U1C, as a mediator of alternative splicing regulation.
authors.affiliation: Institute of Biochemistry, Justus Liebig University of Giessen, Giessen, Germany.
authors.forename: Tanja Dorothe
authors.initials: TD
authors.lastname: Rösel
authors.forename: Lee-Hsueh
authors.initials: LH
authors.lastname: Hung
authors.forename: Jan
authors.initials: J
authors.lastname: Medenbach
authors.forename: Katrin
authors.initials: K
authors.lastname: Donde
authors.forename: Stefan
authors.initials: S
authors.lastname: Starke
authors.forename: Vladimir
authors.initials: V
authors.lastname: Benes
authors.forename: Gunnar
authors.initials: G
authors.lastname: Rätsch
authors.forename: Albrecht
authors.initials: A
authors.lastname: Bindereif
mesh_terms: D017398:Alternative Splicing; D000818:Animals; D004625:Embryo, Nonmammalian / Q000502:physiology; D005786:Gene Expression Regulation; D005816:Genetic Complementation Test; D006367:HeLa Cells; D006801:Humans; D008969:Molecular Sequence Data; D009154:Mutation; D012322:RNA Precursors / Q000737:chemistry / Q000378:metabolism; D017411:Ribonucleoproteins, Small Nuclear / Q000235:genetics / Q000378:metabolism*; D017422:Sequence Analysis, DNA; D015027:Zebrafish / Q000235:genetics / Q000502:physiology*; D029961:Zebrafish Proteins / Q000235:genetics / Q000378:metabolism*
chemical_list: D012322:RNA Precursors; D017411:Ribonucleoproteins, Small Nuclear; D029961:Zebrafish Proteins