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PMID: 16332694 Published · epublish English Journal Article Research Support, Non-U.S. Gov't

SNEV is an evolutionarily conserved splicing factor whose oligomerization is necessary for spliceosome assembly.

Nucleic acids research ·Vol. 33 ·No. 21 ·2005-00-00 ·Pages 6868-83

Grillari J, Ajuh P, Stadler G, Löscher M, Voglauer R, Ernst W, Chusainow J, Eisenhaber F, Pokar M, Fortschegger K, Grey M, Lamond AI, Katinger H

Abstract

We have isolated the human protein SNEV as downregulated in replicatively senescent cells. Sequence homology to the yeast splicing factor Prp19 suggested that SNEV might be the orthologue of Prp19 and therefore might also be involved in pre-mRNA splicing. We have used various approaches including gene complementation studies in yeast using a temperature sensitive mutant with a pleiotropic phenotype and SNEV immunodepletion from human HeLa nuclear extracts to determine its function. A human-yeast chimera was indeed capable of restoring the wild-type phenotype of the yeast mutant strain. In addition, immunodepletion of SNEV from human nuclear extracts resulted in a decrease of in vitro pre-mRNA splicing efficiency. Furthermore, as part of our analysis of protein-protein interactions within the CDC5L complex, we found that SNEV interacts with itself. The self-interaction domain was mapped to amino acids 56-74 in the protein's sequence and synthetic peptides derived from this region inhibit in vitro splicing by surprisingly interfering with spliceosome formation and stability. These results indicate that SNEV is the human orthologue of yeast PRP19, functions in splicing and that homo-oligomerization of SNEV in HeLa nuclear extract is essential for spliceosome assembly and that it might also be important for spliceosome stability.

MeSH Terms
Amino Acid Sequence Cell Nucleus/chemistry Conserved Sequence DNA Repair Enzymes Evolution, Molecular HeLa Cells Humans Molecular Sequence Data Mutation Nuclear Proteins Peptides/pharmacology Phenotype Protein Structure, Tertiary RNA Precursors/metabolism RNA Splicing/drug effects RNA Splicing Factors RNA, Messenger/metabolism RNA-Binding Proteins/analysis Recombinant Proteins/metabolism Saccharomyces cerevisiae Proteins/chemistry,genetics Sequence Homology, Amino Acid Spliceosomes/drug effects,metabolism Ubiquitin-Protein Ligases/chemistry,genetics,physiology
Chemicals
Nuclear Proteins PRP19 protein, S cerevisiae Peptides RNA Precursors RNA Splicing Factors RNA, Messenger RNA-Binding Proteins Recombinant Proteins Saccharomyces cerevisiae Proteins Ubiquitin-Protein Ligases DNA Repair Enzymes PRPF19 protein, human
Authors & Affiliations
13 authors, click to expand affiliations / ORCID
Grillari Johannes
Institute of Applied Microbiology, University of Natural Resources and Applied Life Sciences Vienna, Austria Muthgasse 18, A-1190 Vienna. j.grillari@iam.boku.ac.at
Ajuh Paul
Stadler Guido
Löscher Marlies
Voglauer Regina
Ernst Wolfgang
Chusainow Janet
Eisenhaber Frank
Pokar Marion
Fortschegger Klaus
Grey Martin
Lamond Angus I
Katinger Hermann
References (54)
54 references, click to expand
  1. Characterization of interactions among the Cef1p-Prp19p-associated splicing complex.
    RNA. 2002 Jun;8(6):798-815 PMID: 12088152
  2. Functional analysis of the human CDC5L complex and identification of its components by mass spectrometry.
    EMBO J. 2000 Dec 1;19(23):6569-81 PMID: 11101529
  3. Small nuclear ribonucleoprotein remodeling during catalytic activation of the spliceosome.
    Science. 2002 Dec 13;298(5601):2205-8 PMID: 12411573
  4. Protein composition of human prespliceosomes isolated by a tobramycin affinity-selection method.
    Proc Natl Acad Sci U S A. 2002 Dec 24;99(26):16719-24 PMID: 12477934
  5. Structural insights into the U-box, a domain associated with multi-ubiquitination.
    Nat Struct Biol. 2003 Apr;10(4):250-5 PMID: 12627222
  6. Identification and characterization of two novel components of the Prp19p-associated complex, Ntc30p and Ntc20p.
    J Biol Chem. 2001 Jan 5;276(1):488-94 PMID: 11018040
  7. In vivo analysis of Cajal body movement, separation, and joining in live human cells.
    J Cell Biol. 2000 Dec 25;151(7):1561-74 PMID: 11134083
  8. The F-box protein family.
    Genome Biol. 2000;1(5):REVIEWS3002 PMID: 11178263
  9. Themes and variations on ubiquitylation.
    Nat Rev Mol Cell Biol. 2001 Mar;2(3):169-78 PMID: 11265246
  10. U box proteins as a new family of ubiquitin-protein ligases.
    J Biol Chem. 2001 Aug 31;276(35):33111-20 PMID: 11435423
  11. The pso mutants of Saccharomyces cerevisiae comprise two groups: one deficient in DNA repair and another with altered mutagen metabolism.
    Mutat Res. 2001 Oct;489(1):79-96 PMID: 11673090
  12. A direct interaction between the carboxyl-terminal region of CDC5L and the WD40 domain of PLRG1 is essential for pre-mRNA splicing.
    J Biol Chem. 2001 Nov 9;276(45):42370-81 PMID: 11544257
  13. Snt309p modulates interactions of Prp19p with its associated components to stabilize the Prp19p-associated complex essential for pre-mRNA splicing.
    Proc Natl Acad Sci U S A. 1999 May 11;96(10):5406-11 PMID: 10318896
  14. Role of human Pso4 in mammalian DNA repair and association with terminal deoxynucleotidyl transferase.
    Proc Natl Acad Sci U S A. 2003 Sep 16;100(19):10746-51 PMID: 12960389
  15. The Prp19p-associated complex in spliceosome activation.
    Science. 2003 Oct 10;302(5643):279-82 PMID: 12970570
  16. Identification of peptide inhibitors of pre-mRNA splicing derived from the essential interaction domains of CDC5L and PLRG1.
    Nucleic Acids Res. 2003 Nov 1;31(21):6104-16 PMID: 14576297
  17. A subset of human 35S U5 proteins, including Prp19, function prior to catalytic step 1 of splicing.
    EMBO J. 2004 Jun 16;23(12):2381-91 PMID: 15175653
  18. Cleavage of structural proteins during the assembly of the head of bacteriophage T4.
    Nature. 1970 Aug 15;227(5259):680-5 PMID: 5432063
  19. Isolation and characterization of pso mutants sensitive to photo-addition of psoralen derivatives in Saccharomyces cerevisiae.
    Genetics. 1980 Jun;95(2):273-88 PMID: 7009316
  20. The structure of mammalian small nuclear ribonucleoproteins. Identification of multiple protein components reactive with anti-(U1)ribonucleoprotein and anti-Sm autoantibodies.
    J Biol Chem. 1984 May 10;259(9):5907-14 PMID: 6232278
  21. Electrophoretic separation of complexes involved in the splicing of precursors to mRNAs.
    Cell. 1986 Sep 12;46(6):845-55 PMID: 2944598
  22. Interactions between small nuclear ribonucleoprotein particles in formation of spliceosomes.
    Cell. 1987 Jun 19;49(6):763-74 PMID: 2953438
  23. A mutational analysis of spliceosome assembly: evidence for splice site collaboration during spliceosome formation.
    Genes Dev. 1987 Aug;1(6):532-43 PMID: 2824284
  24. PSO4: a novel gene involved in error-prone repair in Saccharomyces cerevisiae.
    Mutat Res. 1989 Sep;218(2):111-24 PMID: 2671705
  25. Methods and algorithms for statistical analysis of protein sequences.
    Proc Natl Acad Sci U S A. 1992 Mar 15;89(6):2002-6 PMID: 1549558
  26. Improved method for high efficiency transformation of intact yeast cells.
    Nucleic Acids Res. 1992 Mar 25;20(6):1425 PMID: 1561104
  27. The WD-40 repeat.
    FEBS Lett. 1992 Jul 28;307(2):131-4 PMID: 1644165
  28. Yeast precursor mRNA processing protein PRP19 associates with the spliceosome concomitant with or just after dissociation of U4 small nuclear RNA.
    Proc Natl Acad Sci U S A. 1993 Nov 15;90(22):10821-5 PMID: 8248176
  29. Functional association of essential splicing factor(s) with PRP19 in a protein complex.
    EMBO J. 1994 May 15;13(10):2421-31 PMID: 8194532
  30. TopPred II: an improved software for membrane protein structure predictions.
    Comput Appl Biosci. 1994 Dec;10(6):685-6 PMID: 7704669
  31. Further characterization of the yeast pso4-1 mutant: interaction with rad51 and rad52 mutants after photoinduced psoralen lesions.
    Curr Genet. 1996 Feb;29(3):211-8 PMID: 8595666
  32. Analysis of compositionally biased regions in sequence databases.
    Methods Enzymol. 1996;266:554-71 PMID: 8743706
  33. Allelism of PSO4 and PRP19 links pre-mRNA processing with recombination and error-prone DNA repair in Saccharomyces cerevisiae.
    Nucleic Acids Res. 1996 Oct 15;24(20):4009-14 PMID: 8918805
  34. Alpha-helical coiled-coil oligomerization domains in extracellular proteins.
    Matrix Biol. 1997 Mar;15(8-9):555-65; discussion 567-8 PMID: 9138288
  35. Protein functions in pre-mRNA splicing.
    Curr Opin Cell Biol. 1997 Jun;9(3):320-8 PMID: 9159080
  36. Predicting coiled-coil regions in proteins.
    Curr Opin Struct Biol. 1997 Jun;7(3):388-93 PMID: 9204281
  37. Gapped BLAST and PSI-BLAST: a new generation of protein database search programs.
    Nucleic Acids Res. 1997 Sep 1;25(17):3389-402 PMID: 9254694
  38. Mechanical devices of the spliceosome: motors, clocks, springs, and things.
    Cell. 1998 Feb 6;92(3):315-26 PMID: 9476892
  39. The ubiquitin-proteasome pathway: the complexity and myriad functions of proteins death.
    Proc Natl Acad Sci U S A. 1998 Mar 17;95(6):2727-30 PMID: 9501156
  40. Snt309p, a component of the Prp19p-associated complex that interacts with Prp19p and associates with the spliceosome simultaneously with or immediately after dissociation of U4 in the same manner as Prp19p.
    Mol Cell Biol. 1998 Apr;18(4):2196-204 PMID: 9528791
  41. A novel ubiquitination factor, E4, is involved in multiubiquitin chain assembly.
    Cell. 1999 Mar 5;96(5):635-44 PMID: 10089879
  42. Cef1p is a component of the Prp19p-associated complex and essential for pre-mRNA splicing.
    J Biol Chem. 1999 Apr 2;274(14):9455-62 PMID: 10092627
  43. Structural and functional analysis of essential pre-mRNA splicing factor Prp19p.
    Mol Cell Biol. 2005 Jan;25(1):451-60 PMID: 15601865
  44. Interaction of U-box E3 ligase SNEV with PSMB4, the beta7 subunit of the 20 S proteasome.
    Biochem J. 2005 Jun 1;388(Pt 2):593-603 PMID: 15660529
  45. Cotranscriptional spliceosome assembly occurs in a stepwise fashion and requires the cap binding complex.
    Mol Cell. 2005 Jul 1;19(1):53-63 PMID: 15989964
  46. The Isy1p component of the NineTeen complex interacts with the ATPase Prp16p to regulate the fidelity of pre-mRNA splicing.
    Genes Dev. 2005 Aug 15;19(16):1894-904 PMID: 16103217
  47. SNEV overexpression extends the life span of human endothelial cells.
    Exp Cell Res. 2006 Apr 1;312(6):746-59 PMID: 16388800
  48. SMART: a web-based tool for the study of genetically mobile domains.
    Nucleic Acids Res. 2000 Jan 1;28(1):231-4 PMID: 10592234
  49. The Pfam protein families database.
    Nucleic Acids Res. 2000 Jan 1;28(1):263-6 PMID: 10592242
  50. The U box is a modified RING finger - a common domain in ubiquitination.
    Curr Biol. 2000 Feb 24;10(4):R132-4 PMID: 10704423
  51. Subtractive hybridization of mRNA from early passage and senescent endothelial cells.
    Exp Gerontol. 2000 Mar;35(2):187-97 PMID: 10767578
  52. Homology-based method for identification of protein repeats using statistical significance estimates.
    J Mol Biol. 2000 May 5;298(3):521-37 PMID: 10772867
  53. hNMP 200: a novel human common nuclear matrix protein combining structural and regulatory functions.
    Exp Cell Res. 2000 Nov 25;261(1):166-79 PMID: 11082287
  54. Thermoconditional modulation of the pleiotropic sensitivity phenotype by the Saccharomyces cerevisiae PRP19 mutant allele pso4-1.
    Nucleic Acids Res. 2002 Nov 15;30(22):4993-5003 PMID: 12434004
Article Info
Journal
Nucleic acids research
Abbr.
Nucleic Acids Res
ISSN
1362-4962
DOI
10.1093/nar/gki986
Email
j.grillari@iam.boku.ac.at
Published
2005-00-00
Epub
2005-00-06
Pages
6868-83
Language
English
Region
England
NLM ID
0411011
PMCID
PMC1310963
Subset
IM
Grants
Wellcome Trust · 073980 · United Kingdom
External Links
PubMed source DOI: 10.1093/nar/gki986 PMC Full Text View journal details
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