Home LiteratureArticle Details
PMID: 7664739 Published · ppublish English Journal Article

SLU7 and a novel activity, SSF1, act during the PRP16-dependent step of yeast pre-mRNA splicing.

The EMBO journal ·Vol. 14 ·No. 16 ·1995-08-15 ·Pages 4001-9

Ansari A, Schwer B

Abstract

Understanding the mechanism of pre-mRNA splicing requires the characterization of all components involved. In the present study, we used the genetically and biochemically defined yeast PRP16 protein as a point of departure for the identification of additional factors required for the second catalytic step in vitro. We isolated by glycerol gradient sedimentation spliceosomes that were formed in yeast extracts depleted of PRP16. This procedure separated the spliceosomal complexes containing lariat intermediate and exon 1 from free proteins present in the whole-cell yeast extract. We then supplemented these spliceosomes with purified proteins or yeast extract fractions as a functional assay for second-step splicing factors. We show that SLU7 protein and a novel activity that we named SSF1 (second-step factor 1) were required in concert with PRP16 to promote progression through the second catalytic step of splicing. Taking advantage of a differential ATP requirement for PRP16 and SLU7 function, we show that SLU7 can act after PRP16 in the splicing pathway.

MeSH Terms
Actins/genetics Adenosine Triphosphatases/isolation & purification,physiology Adenosine Triphosphate/metabolism Amino Acid Sequence Cell Fractionation Fungal Proteins/genetics,isolation & purification,physiology Genes, Fungal/genetics Molecular Sequence Data RNA Helicases RNA Precursors/genetics RNA Splicing/physiology RNA Splicing Factors RNA, Fungal/genetics RNA, Messenger/genetics RNA-Binding Proteins/metabolism Recombinant Fusion Proteins/isolation & purification,physiology Ribonucleoproteins, Small Nuclear Saccharomyces cerevisiae/genetics Saccharomyces cerevisiae Proteins Spliceosomes/metabolism
Chemicals
Actins Fungal Proteins RNA Precursors RNA Splicing Factors RNA, Fungal RNA, Messenger RNA-Binding Proteins Recombinant Fusion Proteins Ribonucleoproteins, Small Nuclear SLU7 protein, S cerevisiae Saccharomyces cerevisiae Proteins Adenosine Triphosphate Adenosine Triphosphatases PRP16 protein, S cerevisiae RNA Helicases
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Ansari A
Department of Biochemistry, Robert Wood Johnson Medical School, Piscataway, NJ 08854, USA.
Schwer B
References (33)
33 references, click to expand
  1. RNA splicing products formed with isolated fractions from HeLa cells are associated with fast-sedimenting complexes.
    Proc Natl Acad Sci U S A. 1986 Feb;83(4):887-91 PMID: 3456573
  2. Yeast mRNA splicing in vitro.
    J Biol Chem. 1985 Nov 25;260(27):14780-92 PMID: 2997224
  3. An ordered pathway of snRNP binding during mammalian pre-mRNA splicing complex assembly.
    EMBO J. 1987 Aug;6(8):2415-24 PMID: 2959470
  4. Splicing of yeast nuclear pre-mRNA in vitro requires a functional 40S spliceosome and several extrinsic factors.
    Genes Dev. 1987 Mar;1(1):7-18 PMID: 3322937
  5. Spliceosome assembly in yeast.
    Genes Dev. 1987 Nov;1(9):1014-27 PMID: 2962902
  6. Spliceosomal snRNAs.
    Annu Rev Genet. 1988;22:387-419 PMID: 2977088
  7. Isolation and characterization of pre-mRNA splicing mutants of Saccharomyces cerevisiae.
    Genes Dev. 1989 Aug;3(8):1206-16 PMID: 2676722
  8. PRP16 is an RNA-dependent ATPase that interacts transiently with the spliceosome.
    Nature. 1991 Feb 7;349(6309):494-9 PMID: 1825134
  9. Tackling the protease problem in Saccharomyces cerevisiae.
    Methods Enzymol. 1991;194:428-53 PMID: 2005802
  10. Pre-mRNA splicing in yeast.
    Trends Genet. 1991 Mar;7(3):79-85 PMID: 2031287
  11. Messenger RNA splicing in yeast: clues to why the spliceosome is a ribonucleoprotein.
    Science. 1991 Jul 12;253(5016):157-63 PMID: 1853200
  12. Alterations of RNase H sensitivity of the 3' splice site region during the in vitro splicing reaction.
    Nucleic Acids Res. 1991 Jul 25;19(14):3953-8 PMID: 1650457
  13. The purified yeast pre-mRNA splicing factor PRP2 is an RNA-dependent NTPase.
    EMBO J. 1992 Jun;11(6):2319-26 PMID: 1534753
  14. A dominant negative mutation in a spliceosomal ATPase affects ATP hydrolysis but not binding to the spliceosome.
    Mol Cell Biol. 1992 Aug;12(8):3540-7 PMID: 1385854
  15. Roles of PRP8 protein in the assembly of splicing complexes.
    EMBO J. 1992 Oct;11(10):3721-9 PMID: 1396567
  16. 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
  17. An essential splicing factor, SLU7, mediates 3' splice site choice in yeast.
    Genes Dev. 1992 Nov;6(11):2112-24 PMID: 1427075
  18. A conformational rearrangement in the spliceosome is dependent on PRP16 and ATP hydrolysis.
    EMBO J. 1992 Dec;11(13):5033-9 PMID: 1464325
  19. Stages in the second reaction of pre-mRNA splicing: the final step is ATP independent.
    Genes Dev. 1993 Feb;7(2):320-9 PMID: 8436300
  20. A U5 small nuclear ribonucleoprotein particle protein involved only in the second step of pre-mRNA splicing in Saccharomyces cerevisiae.
    Mol Cell Biol. 1993 May;13(5):2959-70 PMID: 8474454
  21. Non-snRNP protein splicing factors.
    Biochim Biophys Acta. 1993 Jun 25;1173(3):247-65 PMID: 8318535
  22. Mammalian protein factors involved in nuclear pre-mRNA splicing.
    Mol Biol Rep. 1993 Aug;18(2):93-8 PMID: 8232300
  23. A dominant negative mutation in the conserved RNA helicase motif 'SAT' causes splicing factor PRP2 to stall in spliceosomes.
    EMBO J. 1994 Feb 15;13(4):879-87 PMID: 8112301
  24. A novel set of spliceosome-associated proteins and the essential splicing factor PSF bind stably to pre-mRNA prior to catalytic step II of the splicing reaction.
    EMBO J. 1994 Jul 15;13(14):3356-67 PMID: 8045264
  25. Interaction of the yeast splicing factor PRP8 with substrate RNA during both steps of splicing.
    Nucleic Acids Res. 1995 Feb 11;23(3):320-6 PMID: 7885825
  26. Dynamic RNA-RNA interactions in the spliceosome.
    Annu Rev Genet. 1994;28:1-26 PMID: 7534458
  27. A novel role for a U5 snRNP protein in 3' splice site selection.
    Genes Dev. 1995 Apr 1;9(7):855-68 PMID: 7535718
  28. The "spliceosome": yeast pre-messenger RNA associates with a 40S complex in a splicing-dependent reaction.
    Science. 1985 May 24;228(4702):963-7 PMID: 3890181
  29. An RNA processing activity that debranches RNA lariats.
    Science. 1985 Jul 12;229(4709):135-40 PMID: 2990042
  30. A multicomponent complex is involved in the splicing of messenger RNA precursors.
    Cell. 1985 Aug;42(1):345-53 PMID: 3160482
  31. Stepwise assembly of a pre-mRNA splicing complex requires U-snRNPs and specific intron sequences.
    Cell. 1985 Aug;42(1):355-67 PMID: 3160483
  32. Multiple factors including the small nuclear ribonucleoproteins U1 and U2 are necessary for pre-mRNA splicing in vitro.
    Cell. 1985 Oct;42(3):725-36 PMID: 2996774
  33. Use of bacteriophage T7 RNA polymerase to direct selective high-level expression of cloned genes.
    J Mol Biol. 1986 May 5;189(1):113-30 PMID: 3537305
Article Info
Journal
The EMBO journal
Abbr.
EMBO J
ISSN
0261-4189
Published
1995-08-15
Pages
4001-9
Language
English
Region
England
NLM ID
8208664
PMCID
PMC394478
Subset
IM
External Links
PubMed source 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: product@genelibs.com

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