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PMID: 9528778 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Synthetic lethality of yeast slt mutations with U2 small nuclear RNA mutations suggests functional interactions between U2 and U5 snRNPs that are important for both steps of pre-mRNA splicing.

Molecular and cellular biology ·Vol. 18 ·No. 4 ·1998-04-00 ·Pages 2055-66

Xu D, Field DJ, Tang SJ, Moris A, Bobechko BP, Friesen JD

Abstract

A genetic screen was devised to identify Saccharomyces cerevisiae splicing factors that are important for the function of the 5' end of U2 snRNA. Six slt (stands for synthetic lethality with U2) mutants were isolated on the basis of synthetic lethality with a U2 snRNA mutation that perturbs the U2-U6 snRNA helix II interaction. SLT11 encodes a new splicing factor and SLT22 encodes a new RNA-dependent ATPase RNA helicase (D. Xu, S. Nouraini, D. Field, S. J. Tang, and J. D. Friesen, Nature 381:709-713, 1996). The remaining four slt mutations are new alleles of previously identified splicing genes: slt15, previously identified as prp17 (slt15/prp17-100), slt16/smd3-1, slt17/slu7-100, and slt21/prp8-21. slt11-1 and slt22-1 are synthetically lethal with mutations in the 3' end of U6 snRNA, a region that affects U2-U6 snRNA helix II; however, slt17/slu7-100 and slt21/prp8-21 are not. This difference suggests that the latter two factors are unlikely to be involved in interactions with U2-U6 snRNA helix II but rather are specific to interactions with U2 snRNA. Pairwise synthetic lethality was observed among slt11-1 (which affects the first step of splicing) and several second-step factors, including slt15/prp17-100, slt17/slu7-100, and prp16-1. Mutations in loop 1 of U5 snRNA, a region that is implicated in the alignment of the two exons, are synthetically lethal with slu4/prp17-2 and slu7-1 (D. Frank, B. Patterson, and C. Guthrie, Mol. Cell. Biol. 12:5179-5205, 1992), as well as with slt11-1, slt15/prp17-100, slt17/slu7-100, and slt21/prp8-21. These same U5 snRNA mutations also interact genetically with certain U2 snRNA mutations that lie in the helix I and helix II regions of the U2-U6 snRNA structure. Our results suggest interactions among U2 snRNA, U5 snRNA, and Slt protein factors that may be responsible for coupling and coordination of the two reactions of pre-mRNA splicing.

MeSH Terms
Base Sequence Cell Cycle Proteins DNA-Binding Proteins Fungal Proteins/genetics Genes, Lethal Molecular Sequence Data Mutation Nucleic Acid Conformation Phenotype RNA Precursors/genetics,physiology RNA Splicing/genetics,physiology RNA Splicing Factors RNA, Small Nuclear/genetics,physiology RNA-Binding Proteins Ribonucleoprotein, U2 Small Nuclear/genetics,physiology Ribonucleoprotein, U4-U6 Small Nuclear Ribonucleoprotein, U5 Small Nuclear/genetics,physiology Ribonucleoproteins, Small Nuclear/genetics Saccharomyces cerevisiae/genetics Saccharomyces cerevisiae Proteins
Chemicals
CDC40 protein, S cerevisiae Cell Cycle Proteins DNA-Binding Proteins Fungal Proteins PRP8 protein, S cerevisiae RNA Precursors RNA Splicing Factors RNA, Small Nuclear RNA-Binding Proteins Ribonucleoprotein, U2 Small Nuclear Ribonucleoprotein, U4-U6 Small Nuclear Ribonucleoprotein, U5 Small Nuclear Ribonucleoproteins, Small Nuclear SLU7 protein, S cerevisiae SMD3 protein, S cerevisiae Saccharomyces cerevisiae Proteins
Authors & Affiliations
6 authors, click to expand affiliations / ORCID
Xu D
Banting and Best Department of Medical Research, University of Toronto, Ontario, Canada.
Field D J
Tang S J
Moris A
Bobechko B P
Friesen J D
References (52)
52 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. CUS1, a suppressor of cold-sensitive U2 snRNA mutations, is a novel yeast splicing factor homologous to human SAP 145.
    Genes Dev. 1996 Jan 15;10(2):220-32 PMID: 8566755
  3. A conformational rearrangement in the spliceosome is dependent on PRP16 and ATP hydrolysis.
    EMBO J. 1992 Dec;11(13):5033-9 PMID: 1464325
  4. Evidence for an essential non-Watson-Crick interaction between the first and last nucleotides of a nuclear pre-mRNA intron.
    Nature. 1993 Feb 18;361(6413):660-2 PMID: 8437627
  5. A mechanism to enhance mRNA splicing fidelity: the RNA-dependent ATPase Prp16 governs usage of a discard pathway for aberrant lariat intermediates.
    Cell. 1993 Jul 2;73(7):1377-91 PMID: 8324826
  6. Four yeast spliceosomal proteins (PRP5, PRP9, PRP11, and PRP21) interact to promote U2 snRNP binding to pre-mRNA.
    Genes Dev. 1993 Oct;7(10):1909-25 PMID: 8405998
  7. Site-specific cross-linking of mammalian U5 snRNP to the 5' splice site before the first step of pre-mRNA splicing.
    Genes Dev. 1992 Dec;6(12B):2542-53 PMID: 1340469
  8. The phylogenetically predicted base-pairing interaction between alpha and alpha' is required for group II splicing in vitro.
    Proc Natl Acad Sci U S A. 1993 Nov 15;90(22):10658-62 PMID: 7504276
  9. Mutations in U6 snRNA that alter splice site specificity: implications for the active site.
    Science. 1993 Dec 24;262(5142):1982-8 PMID: 8266093
  10. The U5 and U6 small nuclear RNAs as active site components of the spliceosome.
    Science. 1993 Dec 24;262(5142):1989-96 PMID: 8266094
  11. Involvement of U6 snRNA in 5' splice site selection.
    Science. 1993 Dec 24;262(5142):2035-9 PMID: 8266100
  12. Mutational analysis of the PRP4 protein of Saccharomyces cerevisiae suggests domain structure and snRNP interactions.
    Nucleic Acids Res. 1994 May 11;22(9):1724-34 PMID: 8202378
  13. Branch nucleophile selection in pre-mRNA splicing: evidence for the bulged duplex model.
    Genes Dev. 1994 Mar 1;8(5):587-97 PMID: 7926752
  14. Randomization-selection analysis of snRNAs in vivo: evidence for a tertiary interaction in the spliceosome.
    Genes Dev. 1994 May 1;8(9):1071-86 PMID: 7926788
  15. Dynamic association of proteins with the pre-mRNA branch region.
    Genes Dev. 1994 Dec 15;8(24):3008-20 PMID: 8001820
  16. 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
  17. Dynamic RNA-RNA interactions in the spliceosome.
    Annu Rev Genet. 1994;28:1-26 PMID: 7534458
  18. A novel role for a U5 snRNP protein in 3' splice site selection.
    Genes Dev. 1995 Apr 1;9(7):855-68 PMID: 7535718
  19. Extensive interactions of PRP8 protein with the 5' and 3' splice sites during splicing suggest a role in stabilization of exon alignment by U5 snRNA.
    EMBO J. 1995 Jun 1;14(11):2602-12 PMID: 7781612
  20. SLU7 and a novel activity, SSF1, act during the PRP16-dependent step of yeast pre-mRNA splicing.
    EMBO J. 1995 Aug 15;14(16):4001-9 PMID: 7664739
  21. Characterization and functional ordering of Slu7p and Prp17p during the second step of pre-mRNA splicing in yeast.
    Proc Natl Acad Sci U S A. 1995 Oct 10;92(21):9687-91 PMID: 7568198
  22. An element in human U6 RNA destabilizes the U4/U6 spliceosomal RNA complex.
    RNA. 1995 Apr;1(2):122-31 PMID: 7585242
  23. Branch-point attack in group II introns is a highly reversible transesterification, providing a potential proofreading mechanism for 5'-splice site selection.
    RNA. 1995 Jun;1(4):391-406 PMID: 7493317
  24. Prp16p, Slu7p, and Prp8p interact with the 3' splice site in two distinct stages during the second catalytic step of pre-mRNA splicing.
    RNA. 1995 Aug;1(6):584-97 PMID: 7489518
  25. snRNA interactions at 5' and 3' splice sites monitored by photoactivated crosslinking in yeast spliceosomes.
    RNA. 1995 Nov;1(9):968-80 PMID: 8548661
  26. Functionally redundant interactions between U2 and U6 spliceosomal snRNAs.
    Genes Dev. 1996 Feb 15;10(4):489-501 PMID: 8600031
  27. Invariant U2 RNA sequences bordering the branchpoint recognition region are essential for interaction with yeast SF3a and SF3b subunits.
    Mol Cell Biol. 1996 Mar;16(3):818-28 PMID: 8622683
  28. A U1/U4/U5 snRNP complex induced by a 2'-O-methyl-oligoribonucleotide complementary to U5 snRNA.
    Science. 1996 May 10;272(5263):881-4 PMID: 8629024
  29. An RNA-dependent ATPase associated with U2/U6 snRNAs in pre-mRNA splicing.
    Nature. 1996 Jun 20;381(6584):709-13 PMID: 8649518
  30. Identification of novel genes required for yeast pre-mRNA splicing by means of cold-sensitive mutations.
    Genetics. 1996 May;143(1):67-80 PMID: 8722763
  31. The HeLa 200 kDa U5 snRNP-specific protein and its homologue in Saccharomyces cerevisiae are members of the DEXH-box protein family of putative RNA helicases.
    EMBO J. 1996 Aug 1;15(15):4001-15 PMID: 8670905
  32. Requirement for SLU7 in yeast pre-mRNA splicing is dictated by the distance between the branchpoint and the 3' splice site.
    RNA. 1996 Jul;2(7):707-17 PMID: 8756413
  33. The invariant U5 snRNA loop 1 sequence is dispensable for the first catalytic step of pre-mRNA splicing in yeast.
    Cell. 1996 Aug 23;86(4):679-89 PMID: 8752221
  34. Mutagenesis of the yeast gene PRP8 reveals domains governing the specificity and fidelity of 3' splice site selection.
    Genetics. 1996 Jun;143(2):723-39 PMID: 8725222
  35. Interactions of the yeast U6 RNA with the pre-mRNA branch site.
    RNA. 1996 Nov;2(11):1110-23 PMID: 8903342
  36. Mutations within the yeast U4/U6 snRNP protein Prp4 affect a late stage of spliceosome assembly.
    RNA. 1997 Feb;3(2):197-209 PMID: 9042946
  37. Trans-activation of group II intron splicing by nuclear U5 snRNA.
    Nature. 1997 Mar 27;386(6623):417-20 PMID: 9121561
  38. Evidence that U5 snRNP recognizes the 3' splice site for catalytic step II in mammals.
    EMBO J. 1997 Aug 1;16(15):4746-59 PMID: 9303319
  39. Prp43: An RNA helicase-like factor involved in spliceosome disassembly.
    Proc Natl Acad Sci U S A. 1997 Oct 28;94(22):11798-802 PMID: 9342317
  40. Yeast mRNA splicing in vitro.
    J Biol Chem. 1985 Nov 25;260(27):14780-92 PMID: 2997224
  41. Multiple exon-binding sites in class II self-splicing introns.
    Cell. 1987 Jul 3;50(1):17-29 PMID: 3297351
  42. A Saccharomyces cerevisiae genomic plasmid bank based on a centromere-containing shuttle vector.
    Gene. 1987;60(2-3):237-43 PMID: 3327750
  43. Isolation and characterization of pre-mRNA splicing mutants of Saccharomyces cerevisiae.
    Genes Dev. 1989 Aug;3(8):1206-16 PMID: 2676722
  44. A putative ATP binding protein influences the fidelity of branchpoint recognition in yeast splicing.
    Cell. 1990 Mar 9;60(5):705-17 PMID: 2138057
  45. Two domains of yeast U6 small nuclear RNA required for both steps of nuclear precursor messenger RNA splicing.
    Science. 1990 Oct 19;250(4979):404-9 PMID: 2145630
  46. Splice site selection and role of the lariat in a group II intron.
    J Mol Biol. 1991 Jun 5;219(3):415-28 PMID: 1711123
  47. U5 snRNA interacts with exon sequences at 5' and 3' splice sites.
    Cell. 1992 Feb 21;68(4):743-54 PMID: 1739979
  48. Synthetic lethal mutations suggest interactions between U5 small nuclear RNA and four proteins required for the second step of splicing.
    Mol Cell Biol. 1992 Nov;12(11):5197-205 PMID: 1406691
  49. Interactions of small nuclear RNA's with precursor messenger RNA during in vitro splicing.
    Science. 1992 Sep 25;257(5078):1918-25 PMID: 1411506
  50. 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
  51. Mutational analysis of the yeast U2 snRNA suggests a structural similarity to the catalytic core of group I introns.
    Cell. 1992 Nov 27;71(5):819-31 PMID: 1423632
  52. 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.18.4.2055
Published
1998-04-00
Pages
2055-66
Language
English
Region
United States
NLM ID
8109087
PMCID
PMC121436
Subset
IM
External Links
PubMed source DOI: 10.1128/MCB.18.4.2055 PMC Full Text View journal details
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