Home LiteratureArticle Details
PMID: 1406691 Published · ppublish English Journal Article

Synthetic lethal mutations suggest interactions between U5 small nuclear RNA and four proteins required for the second step of splicing.

Molecular and cellular biology ·Vol. 12 ·No. 11 ·1992-11-00 ·Pages 5197-205

Frank D, Patterson B, Guthrie C

Abstract

To investigate the function of the U5 small nuclear ribonucleoprotein (snRNP) in pre-mRNA splicing, we have screened for factors that genetically interact with Saccharomyces cerevisiae U5 snRNA. We isolated trans-acting mutations that exacerbate the phenotypes of conditional alleles of the U5 snRNA and named these genes SLU, for synergistically lethal with U5 snRNA. SLU1 and SLU2 are essential for the first catalytic step of splicing, while SLU7 and SLU4 (an allele of PRP17 [U. Vijayraghavan, M. Company, and J. Abelson, Genes Dev. 3:1206-1216, 1989]) are required only for the second step of splicing. Furthermore, slu4-1 and slu7-1 are lethal in combination with mutations in PRP16 and PRP18, which also function in the second step, but not with mutations in factors required for the first catalytic step, such as PRP8 and PRP4. We infer from these data that SLU4, SLU7, PRP18, PRP16, and the U5 snRNA interact functionally and that a major role of the U5 snRNP is to coordinate a set of factors that are required for the completion of the second catalytic step of splicing.

Related Genes
PRP SLU SNR7
MeSH Terms
Base Sequence Genes, Fungal Genes, Lethal Genes, Synthetic Molecular Sequence Data Mutagenesis, Site-Directed Oligodeoxyribonucleotides RNA Splicing RNA, Fungal RNA, Small Nuclear/genetics,metabolism Ribonucleoproteins, Small Nuclear/genetics,metabolism Saccharomyces cerevisiae/genetics
Chemicals
Oligodeoxyribonucleotides RNA, Fungal RNA, Small Nuclear Ribonucleoproteins, Small Nuclear
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Frank D
Department of Biochemistry and Biophysics, University of California, San Francisco 94143.
Patterson B
Guthrie C
References (45)
45 references, click to expand
  1. A compensatory base change in human U2 snRNA can suppress a branch site mutation.
    Genes Dev. 1989 Oct;3(10):1545-52 PMID: 2612904
  2. PRP16 is an RNA-dependent ATPase that interacts transiently with the spliceosome.
    Nature. 1991 Feb 7;349(6309):494-9 PMID: 1825134
  3. A compensatory base change in U1 snRNA suppresses a 5' splice site mutation.
    Cell. 1986 Sep 12;46(6):827-35 PMID: 3757028
  4. Antisense probes containing 2-aminoadenosine allow efficient depletion of U5 snRNP from HeLa splicing extracts.
    Nucleic Acids Res. 1991 Jun 25;19(12):3193-8 PMID: 1648201
  5. Interactions between small nuclear ribonucleoprotein particles in formation of spliceosomes.
    Cell. 1987 Jun 19;49(6):763-74 PMID: 2953438
  6. Yeast mRNA splicing in vitro.
    J Biol Chem. 1985 Nov 25;260(27):14780-92 PMID: 2997224
  7. PRP4 (RNA4) from Saccharomyces cerevisiae: its gene product is associated with the U4/U6 small nuclear ribonucleoprotein particle.
    Mol Cell Biol. 1989 Sep;9(9):3698-709 PMID: 2528686
  8. 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
  9. Viability of clathrin heavy-chain-deficient Saccharomyces cerevisiae is compromised by mutations at numerous loci: implications for the suppression hypothesis.
    Mol Cell Biol. 1991 Aug;11(8):3868-78 PMID: 2072897
  10. A yeast actin-binding protein is encoded by SAC6, a gene found by suppression of an actin mutation.
    Science. 1989 Jan 13;243(4888):231-3 PMID: 2643162
  11. 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
  12. Splicing a spliceosomal RNA.
    Nature. 1989 Jan 5;337(6202):14-5 PMID: 2909888
  13. Heterogeneous nuclear ribonucleoproteins: role in RNA splicing.
    Science. 1986 Mar 28;231(4745):1534-9 PMID: 3952495
  14. Spliceosome assembly in yeast.
    Genes Dev. 1987 Nov;1(9):1014-27 PMID: 2962902
  15. 5-Fluoroorotic acid as a selective agent in yeast molecular genetics.
    Methods Enzymol. 1987;154:164-75 PMID: 3323810
  16. Identification and purification of a 62,000-dalton protein that binds specifically to the polypyrimidine tract of introns.
    Genes Dev. 1989 Dec;3(12A):1874-86 PMID: 2533575
  17. A protein associated with small nuclear ribonucleoprotein particles recognizes the 3' splice site of premessenger RNA.
    Cell. 1986 Dec 26;47(6):973-84 PMID: 2946421
  18. Evidence from complementation assays in vitro that U5 snRNP is required for both steps of mRNA splicing.
    EMBO J. 1989 Oct;8(10):3105-12 PMID: 2531074
  19. The yeast RNA gene products are essential for mRNA splicing in vitro.
    Cell. 1986 Dec 26;47(6):953-63 PMID: 3536128
  20. Suppressors of a U4 snRNA mutation define a novel U6 snRNP protein with RNA-binding motifs.
    Genes Dev. 1991 May;5(5):773-85 PMID: 1827420
  21. Pre-mRNA splicing in yeast.
    Trends Genet. 1991 Mar;7(3):79-85 PMID: 2031287
  22. Genetic depletion indicates a late role for U5 snRNP during in vitro spliceosome assembly.
    Nucleic Acids Res. 1991 Jul 25;19(14):3857-60 PMID: 1830649
  23. Mammalian pre-mRNA branch site selection by U2 snRNP involves base pairing.
    Genes Dev. 1989 Oct;3(10):1553-61 PMID: 2558966
  24. Mutations at a Zn(II) finger motif in the yeast eIF-2 beta gene alter ribosomal start-site selection during the scanning process.
    Cell. 1988 Aug 26;54(5):621-32 PMID: 3136928
  25. Spliceosomal snRNAs.
    Annu Rev Genet. 1988;22:387-419 PMID: 2977088
  26. U5 snRNA interacts with exon sequences at 5' and 3' splice sites.
    Cell. 1992 Feb 21;68(4):743-54 PMID: 1739979
  27. Mutations in yeast U5 snRNA alter the specificity of 5' splice-site cleavage.
    Cell. 1991 Apr 5;65(1):115-23 PMID: 2013092
  28. A suppressor of a HIS4 transcriptional defect encodes a protein with homology to the catalytic subunit of protein phosphatases.
    Cell. 1989 Feb 24;56(4):527-37 PMID: 2537149
  29. 20S small nuclear ribonucleoprotein U5 shows a surprisingly complex protein composition.
    Proc Natl Acad Sci U S A. 1989 Aug;86(16):6038-42 PMID: 2527369
  30. A protein that specifically recognizes the 3' splice site of mammalian pre-mRNA introns is associated with a small nuclear ribonucleoprotein.
    Cell. 1986 Dec 5;47(5):755-66 PMID: 2946417
  31. Genetic analysis of the yeast cytoskeleton.
    Annu Rev Genet. 1987;21:259-84 PMID: 3327466
  32. An essential yeast snRNA with a U5-like domain is required for splicing in vivo.
    Cell. 1987 Jun 5;49(5):613-24 PMID: 3555841
  33. PRP4: a protein of the yeast U4/U6 small nuclear ribonucleoprotein particle.
    Mol Cell Biol. 1989 Sep;9(9):3710-9 PMID: 2528687
  34. Suppressors of yeast actin mutations.
    Genetics. 1989 Apr;121(4):659-74 PMID: 2656401
  35. 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
  36. A U-rich tract enhances usage of an alternative 3' splice site in yeast.
    Cell. 1991 Jan 11;64(1):181-7 PMID: 1846089
  37. 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
  38. Domains of yeast U4 spliceosomal RNA required for PRP4 protein binding, snRNP-snRNP interactions, and pre-mRNA splicing in vivo.
    Genes Dev. 1990 Jul;4(7):1185-96 PMID: 2145195
  39. Distinct sets of SEC genes govern transport vesicle formation and fusion early in the secretory pathway.
    Cell. 1990 May 18;61(4):723-33 PMID: 2188733
  40. A factor, U2AF, is required for U2 snRNP binding and splicing complex assembly.
    Cell. 1988 Jan 29;52(2):207-19 PMID: 2963698
  41. Identification of a yeast snRNP protein and detection of snRNP-snRNP interactions.
    Cell. 1987 Dec 24;51(6):1019-26 PMID: 2961458
  42. Dominant suppressors of yeast actin mutations that are reciprocally suppressed.
    Genetics. 1989 Apr;121(4):675-83 PMID: 2656402
  43. U5 small nuclear ribonucleoprotein: RNA structure analysis and ATP-dependent interaction with U4/U6.
    Mol Cell Biol. 1989 Aug;9(8):3350-9 PMID: 2552294
  44. Isolation and characterization of pre-mRNA splicing mutants of Saccharomyces cerevisiae.
    Genes Dev. 1989 Aug;3(8):1206-16 PMID: 2676722
  45. Rapid and efficient site-specific mutagenesis without phenotypic selection.
    Methods Enzymol. 1987;154:367-82 PMID: 3323813
Article Info
Journal
Molecular and cellular biology
Abbr.
Mol Cell Biol
ISSN
0270-7306
DOI
10.1128/mcb.12.11.5197-5205.1992
Published
1992-11-00
Pages
5197-205
Language
English
Region
United States
NLM ID
8109087
PMCID
PMC360453
Subset
IM
External Links
PubMed source DOI: 10.1128/mcb.12.11.5197-5205.1992 PMC Full Text View journal details
Analysis Services
Analysis Services

Contact

No. 2 Wenbo Road, Zhangqiu District, Jinan, Shandong

Qilu Normal University · Genelibs Bioinformatics Lab

750 Shunhua Rd, Jinan

2F, Bldg F, University Science Park

Tel: 0531-88819269

WeChat Official Account

Follow our WeChat subscription account for real-time updates and the latest in medical and biological research.


Business Email

E-mail: [email protected]

Copyright © 2026.Genelibs Jinan Honghui Biotechnology Co., Ltd. 鲁ICP备20007159号-2