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PMID: 7565787 Published · ppublish English Comparative Study Journal Article Research Support, U.S. Gov't, P.H.S.

The amino-terminal domain of yeast U1-70K is necessary and sufficient for function.

Molecular and cellular biology ·Vol. 15 ·No. 11 ·1995-11-00 ·Pages 6341-50

Hilleren PJ, Kao HY, Siliciano PG

Abstract

The Saccharomyces cerevisiae SNP1 gene encodes a protein that shares 30% amino acid identity with the mammalian U1 small nuclear ribonucleoprotein particle protein 70K (U1-70K). We have demonstrated that yeast strains in which the SNP1 gene was disrupted are viable but exhibit greatly increased doubling times and severe temperature sensitivity. Furthermore, snp1-null strains are defective in pre-mRNA splicing. We have tested deletion alleles of SNP1 for their ability to complement these phenotypes. We found that the highly conserved RNA recognition motif consensus domain of Snp1 is not required for complementation of the snp1-null growth or splicing defects nor for the in vivo association with the U1 small nuclear ribonucleoprotein particle. However, the amino-terminal domain of Snp1, less strongly conserved, is necessary and sufficient for complementation.

MeSH Terms
Amino Acid Sequence Base Sequence DNA Primers/chemistry Fungal Proteins/chemistry Genetic Complementation Test Molecular Sequence Data Mutagenesis, Site-Directed RNA Splicing RNA, Small Nuclear/metabolism RNA-Binding Proteins/chemistry Ribonucleoprotein, U1 Small Nuclear/chemistry Saccharomyces cerevisiae/genetics Saccharomyces cerevisiae Proteins Sequence Alignment Sequence Deletion Sequence Homology, Amino Acid Structure-Activity Relationship
Chemicals
DNA Primers Fungal Proteins RNA, Small Nuclear RNA-Binding Proteins Ribonucleoprotein, U1 Small Nuclear SNP1 protein, S cerevisiae Saccharomyces cerevisiae Proteins
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Hilleren P J
Department of Biochemistry, University of Minnesota, Minneapolis 55455, USA.
Kao H Y
Siliciano P G
References (58)
58 references, click to expand
  1. An essential splicing factor, SLU7, mediates 3' splice site choice in yeast.
    Genes Dev. 1992 Nov;6(11):2112-24 PMID: 1427075
  2. A common RNA recognition motif identified within a defined U1 RNA binding domain of the 70K U1 snRNP protein.
    Cell. 1989 Apr 7;57(1):89-101 PMID: 2467746
  3. Convergent transcripts of the yeast PRP38-SMD1 locus encode two essential splicing factors, including the D1 core polypeptide of small nuclear ribonucleoprotein particles.
    Proc Natl Acad Sci U S A. 1993 Feb 1;90(3):848-52 PMID: 8430095
  4. Mutational analysis of pre-mRNA splicing in Saccharomyces cerevisiae using a sensitive new reporter gene, CUP1.
    Genetics. 1993 Apr;133(4):851-63 PMID: 8462846
  5. Thiophosphorylation of U1-70K protein inhibits pre-mRNA splicing.
    Nature. 1993 May 20;363(6426):283-6 PMID: 8387646
  6. hnRNP proteins and the biogenesis of mRNA.
    Annu Rev Biochem. 1993;62:289-321 PMID: 8352591
  7. Functional analysis of pre-mRNA splicing factor SF2/ASF structural domains.
    EMBO J. 1993 Dec;12(12):4715-26 PMID: 8223480
  8. Involvement of U6 snRNA in 5' splice site selection.
    Science. 1993 Dec 24;262(5142):2035-9 PMID: 8266100
  9. Analysis of the RNA-recognition motif and RS and RGG domains: conservation in metazoan pre-mRNA splicing factors.
    Nucleic Acids Res. 1993 Dec 25;21(25):5803-16 PMID: 8290338
  10. Purification of small nuclear ribonucleoprotein particles with antibodies against modified nucleosides of small nuclear RNAs.
    Methods Enzymol. 1990;181:232-57 PMID: 2143251
  11. Characterisation of human and murine snRNP proteins by two-dimensional gel electrophoresis and phosphopeptide analysis of U1-specific 70K protein variants.
    Nucleic Acids Res. 1990 Aug 11;18(15):4427-38 PMID: 2143816
  12. Universally conserved and yeast-specific U1 snRNA sequences are important but not essential for U1 snRNP function.
    Genes Dev. 1990 Oct;4(10):1766-74 PMID: 2147421
  13. High-expression vectors with multiple cloning sites for construction of trpE fusion genes: pATH vectors.
    Methods Enzymol. 1991;194:477-90 PMID: 2005804
  14. Mutations in yeast U5 snRNA alter the specificity of 5' splice-site cleavage.
    Cell. 1991 Apr 5;65(1):115-23 PMID: 2013092
  15. Cloning of a yeast U1 snRNP 70K protein homologue: functional conservation of an RNA-binding domain between humans and yeast.
    EMBO J. 1991 Sep;10(9):2627-34 PMID: 1714384
  16. Cloning and domain structure of the mammalian splicing factor U2AF.
    Nature. 1992 Feb 13;355(6361):609-14 PMID: 1538748
  17. Mutations at the 3' splice site can be suppressed by compensatory base changes in U1 snRNA in fission yeast.
    Cell. 1992 Jun 26;69(7):1159-69 PMID: 1617727
  18. The yeast homolog of the U1 snRNP protein 70K is encoded by the SNP1 gene.
    Nucleic Acids Res. 1992 Aug 11;20(15):4009-13 PMID: 1387202
  19. Interactions of small nuclear RNA's with precursor messenger RNA during in vitro splicing.
    Science. 1992 Sep 25;257(5078):1918-25 PMID: 1411506
  20. A novel base-pairing interaction between U2 and U6 snRNAs suggests a mechanism for the catalytic activation of the spliceosome.
    Cell. 1992 Nov 27;71(5):803-17 PMID: 1423631
  21. Heterogeneous nuclear ribonucleoprotein particles and the pathway of mRNA formation.
    Trends Biochem Sci. 1988 Mar;13(3):86-91 PMID: 3072706
  22. Identification of functional U1 snRNA-pre-mRNA complexes committed to spliceosome assembly and splicing.
    Cell. 1989 Oct 20;59(2):349-58 PMID: 2529976
  23. Direct, sequence-specific binding of the human U1-70K ribonucleoprotein antigen protein to loop I of U1 small nuclear RNA.
    Mol Cell Biol. 1989 Oct;9(10):4179-86 PMID: 2531275
  24. Synthesis of small RNAs using T7 RNA polymerase.
    Methods Enzymol. 1989;180:51-62 PMID: 2482430
  25. Structure and expression of the Drosophila melanogaster gene for the U1 small nuclear ribonucleoprotein particle 70K protein.
    Mol Cell Biol. 1990 Jun;10(6):2492-502 PMID: 1692955
  26. Unexpected flexibility in an evolutionarily conserved protein-RNA interaction: genetic analysis of the Sm binding site.
    EMBO J. 1990 Aug;9(8):2555-61 PMID: 2142451
  27. Protein-protein interactions and 5'-splice-site recognition in mammalian mRNA precursors.
    Nature. 1994 Mar 10;368(6467):119-24 PMID: 8139654
  28. Isolation of S. cerevisiae snRNPs: comparison of U1 and U4/U6.U5 to their human counterparts.
    Science. 1994 Apr 8;264(5156):261-5 PMID: 8146658
  29. The association of the U1-specific 70K and C proteins with U1 snRNPs is mediated in part by common U snRNP proteins.
    EMBO J. 1994 Sep 1;13(17):4113-25 PMID: 8076607
  30. The yeast MUD2 protein: an interaction with PRP11 defines a bridge between commitment complexes and U2 snRNP addition.
    Genes Dev. 1994 Apr 1;8(7):843-54 PMID: 7926772
  31. Crystal structure at 1.92 A resolution of the RNA-binding domain of the U1A spliceosomal protein complexed with an RNA hairpin.
    Nature. 1994 Dec 1;372(6505):432-8 PMID: 7984237
  32. Structurally related but functionally distinct yeast Sm D core small nuclear ribonucleoprotein particle proteins.
    Mol Cell Biol. 1995 Jan;15(1):445-55 PMID: 7799953
  33. Sm and Sm-like proteins belong to a large family: identification of proteins of the U6 as well as the U1, U2, U4 and U5 snRNPs.
    EMBO J. 1995 May 1;14(9):2089-98 PMID: 7744014
  34. Structure and expression of a Xenopus gene encoding an snRNP protein (U1 70K).
    EMBO J. 1988 Dec 20;7(13):4311-21 PMID: 2468488
  35. Sequence of U1 RNA from Drosophila melanogaster: implications for U1 secondary structure and possible involvement in splicing.
    Nucleic Acids Res. 1981 Dec 11;9(23):6351-68 PMID: 6172778
  36. Transformation of intact yeast cells treated with alkali cations.
    J Bacteriol. 1983 Jan;153(1):163-8 PMID: 6336730
  37. Isolation of small nuclear ribonucleoproteins containing U1, U2, U4, U5, and U6 RNAs.
    J Biol Chem. 1983 Feb 25;258(4):2604-13 PMID: 6185498
  38. The U1 small nuclear RNA-protein complex selectively binds a 5' splice site in vitro.
    Cell. 1983 Jun;33(2):509-18 PMID: 6190573
  39. Characterization of U small nuclear RNA-associated proteins.
    J Biol Chem. 1984 Oct 25;259(20):12850-6 PMID: 6208190
  40. Lariat structures are in vivo intermediates in yeast pre-mRNA splicing.
    Cell. 1984 Dec;39(3 Pt 2):611-21 PMID: 6096014
  41. The 3' splice site of pre-messenger RNA is recognized by a small nuclear ribonucleoprotein.
    Science. 1985 Dec 20;230(4732):1344-9 PMID: 2933810
  42. Identification of protein sequence homology by consensus template alignment.
    J Mol Biol. 1986 Mar 20;188(2):233-58 PMID: 3088284
  43. A compensatory base change in U1 snRNA suppresses a 5' splice site mutation.
    Cell. 1986 Sep 12;46(6):827-35 PMID: 3757028
  44. RNA-protein organization of U1, U5 and U4-U6 small nuclear ribonucleoproteins in HeLa cells.
    J Mol Biol. 1986 Jun 5;189(3):519-32 PMID: 2946870
  45. mRNA polyadenylate-binding protein: gene isolation and sequencing and identification of a ribonucleoprotein consensus sequence.
    Mol Cell Biol. 1986 Aug;6(8):2932-43 PMID: 3537727
  46. Electrophoresis of ribonucleoproteins reveals an ordered assembly pathway of yeast splicing complexes.
    Nature. 1986 Nov 27-Dec 3;324(6095):341-5 PMID: 3537805
  47. Purification of the individual snRNPs U1, U2, U5 and U4/U6 from HeLa cells and characterization of their protein constituents.
    EMBO J. 1986 Dec 20;5(13):3509-16 PMID: 2951249
  48. Recognition of the TACTAAC box during mRNA splicing in yeast involves base pairing to the U2-like snRNA.
    Cell. 1987 Apr 24;49(2):229-39 PMID: 3552247
  49. S. cerevisiae U1 RNA is large and has limited primary sequence homology to metazoan U1 snRNA.
    Cell. 1987 Aug 14;50(4):593-602 PMID: 2440584
  50. Saccharomyces cerevisiae has a U1-like small nuclear RNA with unexpected properties.
    Science. 1987 Sep 18;237(4821):1484-7 PMID: 3306922
  51. A human autoimmune protein associated with U1 RNA contains a region of homology that is cross-reactive with retroviral p30gag antigen.
    Cell. 1987 Oct 23;51(2):211-20 PMID: 2959371
  52. In vitro assembly of U1 snRNPs.
    EMBO J. 1987 Nov;6(11):3479-85 PMID: 2962858
  53. Spliceosome assembly in yeast.
    Genes Dev. 1987 Nov;1(9):1014-27 PMID: 2962902
  54. A U1 snRNA:pre-mRNA base pairing interaction is required early in yeast spliceosome assembly but does not uniquely define the 5' cleavage site.
    EMBO J. 1988 Aug;7(8):2533-8 PMID: 3056718
  55. An early hierarchic role of U1 small nuclear ribonucleoprotein in spliceosome assembly.
    Science. 1988 Nov 18;242(4881):1028-35 PMID: 2973660
  56. 5' splice site selection in yeast: genetic alterations in base-pairing with U1 reveal additional requirements.
    Genes Dev. 1988 Oct;2(10):1258-67 PMID: 3060402
  57. Loop I of U1 small nuclear RNA is the only essential RNA sequence for binding of specific U1 small nuclear ribonucleoprotein particle proteins.
    Mol Cell Biol. 1988 Nov;8(11):4787-91 PMID: 2974920
  58. Evidence for a base-pairing interaction between U6 small nuclear RNA and 5' splice site during the splicing reaction in yeast.
    Proc Natl Acad Sci U S A. 1992 Dec 1;89(23):11269-73 PMID: 1333604
Article Info
Journal
Molecular and cellular biology
Abbr.
Mol Cell Biol
ISSN
0270-7306
DOI
10.1128/MCB.15.11.6341
Published
1995-11-00
Pages
6341-50
Language
English
Region
United States
NLM ID
8109087
PMCID
PMC230886
Subset
IM
Grants
NIGMS NIH HHS · GM-44628 · United States
External Links
PubMed source DOI: 10.1128/MCB.15.11.6341 PMC Full Text View journal details
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