title: The human U4/U6 snRNP contains 60 and 90kD proteins that are structurally homologous to the yeast splicing factors Prp4p and Prp3p.
abstract: Immunoaffinity-purified human 25S [U4/U6.U5] tri-snRNPs harbor a set of polypeptides, termed the tri-snRNP proteins, that are not present in Mono Q-purified 20S U5 snRNPs or 10S U4/U6 snRNPs and that are important for tri-snRNP complex formation (Behrens SE, Lührmann R, 1991, Genes & Dev 5:1439-1452). Biochemical and immunological characterization of HeLa [U4/U6.U5] tri-snRNPs led to the identification of two novel proteins with molecular weights of 61 and 63kD that are distinct from the previously described 15.5, 20, 27, 60, and 90kD tri-snRNP proteins. For the initial characterization of tri-snRNP proteins that interact directly with U4/U6 snRNPs, immunoaffinity chromatography with an antibody directed against the 60kD protein was performed. We demonstrate that the 60 and 90kD tri-snRNP proteins specifically associate with the U4/U6 snRNP at salt concentrations where the tri-snRNP complex has dissociated. The primary structures of the 60kD and 90kD proteins were determined by cloning and sequencing their respective cDNAs. The U4/U6-60kD protein possesses a C-terminal WD domain that contains seven WD repeats and thus belongs to the WD-protein family, whose best-characterized members include the Gbeta subunits of heterotrimeric G proteins. A database homology search revealed a significant degree of overall homology (57.8% similarity, 33.9% identity) between the human 60kD protein and the Saccharomyces cerevisiae U4/U6 snRNP protein Prp4p. Two additional, previously undetected WD repeats (with seven in total) were also identified in Prp4p, consistent with the possibility that 60kD/Prp4p, like beta-transducin, may adopt a propeller-like structure. The U4/U6-90kD protein was shown to exhibit significant homology, particularly in its C-terminal half, with the S. cerevisiae splicing factor Prp3p, which also associates with the yeast U4/U6 snRNP. Interestingly, U4/U6-90kD shares short regions of homology with E. coli RNase III, including a region encompassing its double-stranded RNA binding domain. Based on their structural similarity with essential splicing factors in yeast, the human U4/U6-60kD and 90kD proteins are likely also to play important roles in the mammalian splicing process.
authors.affiliation: Institut fur Molekularbiologie und Tumorforschung, Marburg, Germany.
authors.forename: J
authors.initials: J
authors.lastname: Lauber
authors.forename: G
authors.initials: G
authors.lastname: Plessel
authors.forename: S
authors.initials: S
authors.lastname: Prehn
authors.forename: C L
authors.initials: CL
authors.lastname: Will
authors.forename: P
authors.initials: P
authors.lastname: Fabrizio
authors.forename: K
authors.initials: K
authors.lastname: Gröning
authors.forename: W S
authors.initials: WS
authors.lastname: Lane
authors.forename: R
authors.initials: R
authors.lastname: Lührmann
mesh_terms: D000595:Amino Acid Sequence; D000906:Antibodies; D001483:Base Sequence; D002846:Chromatography, Affinity / Q000379:methods; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000302:isolation & purification; D004586:Electrophoresis / Q000379:methods; D006367:HeLa Cells; D006801:Humans; D008969:Molecular Sequence Data; D008970:Molecular Weight; D009687:Nuclear Proteins / Q000737:chemistry; D010455:Peptides; D017346:Protein Serine-Threonine Kinases / Q000737:chemistry*; D000072260:RNA Splicing Factors; D017414:Ribonucleoprotein, U4-U6 Small Nuclear / Q000737:chemistry / Q000235:genetics / Q000276:immunology*; D012441:Saccharomyces cerevisiae / Q000737:chemistry; D029701:Saccharomyces cerevisiae Proteins; D017421:Sequence Analysis; D017386:Sequence Homology, Amino Acid
chemical_list: D000906:Antibodies; D018076:DNA, Complementary; D009687:Nuclear Proteins; C491564:PRP3 protein, S cerevisiae; C108507:PRPF3 protein, human; C489982:PRPF4 protein, human; D010455:Peptides; D000072260:RNA Splicing Factors; D017414:Ribonucleoprotein, U4-U6 Small Nuclear; D029701:Saccharomyces cerevisiae Proteins; C083212:PRP4 protein, S cerevisiae; D017346:Protein Serine-Threonine Kinases