title: U1 small nuclear RNA variants differentially form ribonucleoprotein particles in vitro.
abstract: The U1 small nuclear (sn)RNA participates in splicing of pre-mRNAs by recognizing and binding to 5' splice sites at exon/intron boundaries. U1 snRNAs associate with 5' splice sites in the form of ribonucleoprotein particles (snRNPs) that are comprised of the U1 snRNA and 10 core components, including U1A, U1-70K, U1C and the 'Smith antigen', or Sm, heptamer. The U1 snRNA is highly conserved across a wide range of taxa; however, a number of reports have identified the presence of expressed U1-like snRNAs in multiple species, including humans. While numerous U1-like molecules have been shown to be expressed, it is unclear whether these variant snRNAs have the capacity to form snRNPs and participate in splicing. The purpose of the present study was to further characterize biochemically the ability of previously identified human U1-like variants to form snRNPs and bind to U1 snRNP proteins. A bioinformatics analysis provided support for the existence of multiple expressed variants. In vitro gel shift assays, competition assays, and immunoprecipitations (IPs) revealed that the variants formed high molecular weight assemblies to varying degrees and associated with core U1 snRNP proteins to a lesser extent than the canonical U1 snRNA. Together, these data suggest that the human U1 snRNA variants analyzed here are unable to efficiently bind U1 snRNP proteins. The current work provides additional biochemical insights into the ability of the variants to assemble into snRNPs.
authors.affiliation: Center for RNA Biology and Department of Molecular Genetics and Microbiology, Duke University Medical Center, Durham, NC.
authors.forename: Jason A
authors.initials: JA
authors.lastname: Somarelli
authors.affiliation: Department of Biological Sciences, Florida International University, Miami, FL.
authors.forename: Annia
authors.initials: A
authors.lastname: Mesa
authors.affiliation: Department of Biological Sciences, Florida International University, Miami, FL.
authors.forename: Carol E
authors.initials: CE
authors.lastname: Rodriguez
authors.affiliation: Department of Basic Medical Sciences, University of Arizona, College of Medicine- Phoenix, Phoenix, AZ.
authors.forename: Shalini
authors.initials: S
authors.lastname: Sharma
authors.affiliation: Human and Molecular Genetics, College of Medicine, Florida International University, Miami, FL.
authors.forename: Rene J
authors.initials: RJ
authors.lastname: Herrera
mesh_terms: D020224:Expressed Sequence Tags; D014644:Genetic Variation; D006801:Humans; D011485:Protein Binding; D012342:RNA, Small Nuclear / Q000235:genetics*; D017412:Ribonucleoprotein, U1 Small Nuclear / Q000737:chemistry*
chemical_list: D012342:RNA, Small Nuclear; D017412:Ribonucleoprotein, U1 Small Nuclear; C490688:SNRNP70 protein, human; C405483:U1 small nuclear RNA; C075339:U1A protein
keywords: Sm proteins; Spliceosome; Splicing; U1-70K protein; U1A protein