Abstract
Proteins containing RNP consensus-sequence RNA-binding domains (CS-RBDs) play diverse roles in many aspects of RNA metabolism. Using a PCR strategy, we cloned portions of six newSchizosaccharomyces pombe genes encoding RBD proteins, including a putative homolog of the mammalian splicing factor SAP49. The genomic locus corresponding to a second PCR product, designatedrnp24, was cloned and characterized in detail. Sequence analysis revealed that the Rnp24 protein is highly charged and contains a second RBD with an unusually long Loop-3 sequence. Strains containing a disrupted copy of thernp24 gene display neither loss of viability nor any discernible growth defects under a variety of conditions, suggesting that the function of Rnp24p overlaps with that of another fission yeast protein. Although database searches did not identify proteins that share extensive amino-acid identity with Rnp24p, phylogenetic analysis suggests that its closest relatives are metazoan hnRNP proteins. The lack of an observable phenotype inS. pombe cells lacking Rnp24p is consistent with this classification, since hnRNP proteins in higher cells include several distinct subfamilies with similar sequences and RNA-binding specifities.
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References
Adam SA, Nakagawa TY, Swanson MS, Woodruff T, Dreyfuss G (1986) mRNA polyadenylate-binding protein: gene isolation and sequencing and identification of a ribonucleoprotein consensus sequence. Mol Cell Biol 6:2932–2943
Altschul SF, Gish W, Miller W, Myers EW, Lipman DJ (1990) Basic local alignment search tool. J Mol Biol 215:403–410
Apolinaro E, Nocero M, Jin M, Hoffman CS (1994) Cloning and manipulation of theSchizosaccharomyces pombe his7 + gene as a new selectable marker for molecular genetic studies. Curr Genet 24:491–495
Bandziulis RJ, Swanson MS, Dreyfuss G (1989) RNA-binding proteins as developmental regulators. Genes Dev 32:431–437
Bentley RC, Keene JD (1991) Recognition of U1 and U2 small nuclear RNAs can be altered by a 5-amino-acid segment in the U2 small nuclear ribonucleoprotein particle (snRNP) B″ protein and through interactions with the U2 snRNP-A′ protein. Mol Cell Biol 11:1829–1839
Birney E, Kumar S, Krainer AR (1993) Analysis of the RNA-recognition motif and RS and RGG domains: conservation in metazoan pre-mRNA splicing factors. Nucleic Acids Res 21:5803–5816
Bresch C, Muller G, Egel R (1968) Genes involved in meiosis and sporulation of a yeast. Mol Gen Genet 102:301–306
Burd CG, Dreyfuss G (1994) Conserved structures and diversity of functions of RNA-binding proteins. Science 265:615–621
Burd CG, Matunis EL, Dreyfuss G (1991) The multiple RNA-binding domains of the mRNA poly(A)-binding protein have different RNA-binding activities. Mol Cell Biol 11:3419–3424
Champion-Arnaud P, Reed R (1994) The prespliceosome components SAP 49 and SAP 145 interact in a complex implicated in tethering U2 snRNP to the branch site. Genes Dev 8:1974–1983
Cobianchi R, SenGupta DN, Zmudzka BZ, Wilson SH (1986) Structure of rodent helix-destabilizing protein revealed by cDNA cloning. J Biol Chem 261:3536–3543
Cusick ME (1994) RNP1, a new ribonucleoprotein gene of the yeastSaccharomyces cerevisiae. Nucleic Acids Res 22:869–877
Dreyfuss G, Matunis MJ, Pinol-Roma S, Burd CG (1993) hnRNP proteins and the biogenesis of mRNA. Annu Rev Biochem 62:289–321
Fikes JD, Becker DM, Winston F (1990) Striking conservation of TFIID inSchizosaccharomyces pombe andSaccharomyces cerevisiae. Nature 346:291–294
Higgins DG, Sharp PM (1989) Fast and sensitive multiple sequence alignments on a microcomputer. CABIOS 5:151–153
Hoffman DW, Query CC, Golden BL, White SW, Keene JD (1991) RNA-binding domain of the A protein component of the U1 small nuclear ribonucleoprotein analyzed by NMR spectroscopy is structurally similar to ribosomal protein. Proc Natl Acad Sci USA 88:2495–2499
Kim Y-J, Baker BS (1993) Isolation of RRM-type RNA-binding protein genes and the analysis of their relatedness by using a numerical approach. Mol Cell Biol 13:174–183
Liao X, Brennwald P, Wise JA (1989) Genetic analysis ofSchizosaccharomyces pombe 7SL RNA: a structural motif that includes a conserved tetranucleotide loop is important for function. Proc Natl Acad Sci USA 86:4137–4141
Nagai K, Oubridge C, Jessen TH, Li J, Evans PR (1990) Crystal structure of the RNA-binding domain of the U1 small nuclear ribonucleoprotein A. Nature 348:515–520
Pearson WR, Lipman DJ (1988) Improved tools for biological sequence comparison. Proc Natl Acad Sci USA 85:2444–2448
Porter G, Brennwald P, Wise JA (1990) U1 small nuclear RNA fromSchizosaccharomyces pombe has unique and conserved features and is encoded by an essential single-copy gene. Mol Cell Biol 10:2874–2881
Potashkin J, Naik K, Wentz-Hunter K (1993) U2AF homolog required for splicing in vivo. Science 262:573–575
Query CC, Bentley RC, Keene JD (1989) A common RNA recognition motif identified within a defined U1 RNA-binding domain of the 70 U1 snRNP protein. Cell 57:89–101
Riva S, Morandi C, Tsoulfas P, Pandolfo M, Biamonti G, Merrill B, Williams KR, Multhaup G, Beyreuther K, Werr H, Henrich B, Schafer KP (1986) Mammalian single-stranded DNA-binding protein UPI is derived from the ImRNP core protein A1. EMBO J 5:2267–2273
Sachs AB, Bond WM, Kornberg RD (1986) A single gene from yeast for both nuclear and cytoplasmic polyadenylate-binding protein: domain structure and expression. Cell 45:827–835
Saitou N, Nei M (1987) The neighbor-joining method: a new method for reconstructing phylogenetic trees. Mol Biol Evol 4:406–425
Sambrook J, Fritsch EF, Maniatis T (1989) Molecular cloning: a laboratory manual. Cold Spring Harbor Laboratory, Cold Spring Harbor, New York
Scherly D, Boelens W, Dathan NA, van Venrooij WJ, Mattaj IW (1990) Major determinants of the specificity of interaction between small nuclear ribonucleoproteins U1A and U2B″ and their cognate RNAs. Nature 345:502–506
Selinger DA, Porter GL, Brennwald PJ, Wise JA (1992) The two similarly expressed genes encoding U3 snRNA inSchizosaccharomyces pombe lack introns. Mol Biol Evol 9:297–308
Swanson MS, Nakagawa TY, LeVan K, Dreyfuss G (1987) Primary structure of human nuclear ribonucleoprotein particle C proteins: conservation of sequence and domain structures in heterogeneous nuclear RNA, mRNA and pre-rRNA-binding proteins. Mol Cell Biol 7:1731–1739
Thompson JD, Higgins DG, Gibson TJ (1994) CLUSTALW: improving the sensitivity of progressive multiple sequence alignments through sequence weighting, position-specific gap penalties and weight matrix choice. Nucleic Acids Res 22:4673–4680.
Toda T, Shimanuki M, Yanagida M (1991) Fission-yeast genes that confer resistance to staurosporine encode an AP-1-like transcription factor and a protein kinase related to the mammalian ERK1/MAP2 and budding-yeast FUS3 and KSSI kinases. Genes Dev 5:60–73
Toyama R, Goldstein DJ, Schlegel R, Dhar R (1992) A genomic sequence of theSchizosaccharomyces pombe 16-kDa vacuolar H + -ATPase. Yeast 7:989–991
Watanabe Y, Yamamoto M (1994)S. pombe mei2 + encodes an RNA-binding protein essential for premeiotic DNA synthesis and meiosis I, which cooperates with a novel RNA species meiRNA. Cell 78:487–498
Wittekind M, Gorlach M, Friedrichs M, Dreyfuss G, Mueller L (1992) 1H, 13C, and 15N NMR assignments and global folding pattern of the RNA-binding domain of the human hnRNP C proteins. Biochemistry 31:6254–6265
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Communicated by A. Goffeau
GenBank Accession Number U24430
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VanHoy, R.W., Wise, J.A. Molecular analysis of a novelSchizosaccharomyces pombe gene containing two RNP consensus-sequence RNA-binding domains. Curr Genet 29, 307–315 (1996). https://doi.org/10.1007/BF02208611
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DOI: https://doi.org/10.1007/BF02208611