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PMID: 11842115 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Functional and physical interactions between components of the Prp19p-associated complex.

Nucleic acids research ·Vol. 30 ·No. 4 ·2002-02-15 ·Pages 1029-37

Chen CH, Yu WC, Tsao TY, Wang LY, Chen HR, Lin JY, Tsai WY, Cheng SC

Abstract

The Prp19p-associated complex is essential for the yeast pre-mRNA splicing reaction. The complex consists of at least eight protein components, but is not tightly associated with spliceosomal snRNAs. By a combination of genetic and biochemical methods we previously identified four components of this complex, Ntc25p, Ntc85p, Ntc30p and Ntc20p, all of them being novel splicing factors. We have now identified three other components of the complex, Ntc90p, Ntc77p and Ntc31p. These three proteins were also associated with the spliceosome during the splicing reaction in the same manner as Prp19p, concurrently with or immediately after dissociation of U4 snRNA. Two-hybrid analysis revealed that none of these proteins interacted with Prp19p or Ntc25p, but all interacted with Ntc85p. An interaction network between the identified components of the Prp19p-associated complex is demonstrated. Biochemical analysis revealed that Ntc90p, Ntc31p, Ntc30p and Ntc20p form a subcomplex, which, through interacting with Ntc85p and Ntc77p, can associate with Prp19p and Ntc25p to form the Prp19p-associated complex. Genetic analysis suggests that Ntc31p, Ntc30p and Ntc20p may play roles in modulating the function of Ntc90p.

MeSH Terms
Cell Cycle Proteins/genetics,metabolism,physiology Cell Division Fungal Proteins/genetics,metabolism,physiology Macromolecular Substances Models, Biological Mutation Precipitin Tests RNA Splicing RNA Splicing Factors RNA, Fungal/metabolism Saccharomyces cerevisiae/growth & development,metabolism Saccharomyces cerevisiae Proteins Spliceosomes/metabolism Two-Hybrid System Techniques
Chemicals
CLF1 protein, S cerevisiae Cell Cycle Proteins Fungal Proteins Macromolecular Substances PRP19 protein, S cerevisiae RNA Splicing Factors RNA, Fungal Saccharomyces cerevisiae Proteins Syf1 protein, S cerevisiae
Authors & Affiliations
8 authors, click to expand affiliations / ORCID
Chen Chun-Hong
Institute of Molecular Biology, Academia Sinica, Nankang, Taiwan, Republic of China.
Yu Wan-Chin
Tsao Twee Y
Wang Lian-Yung
Chen Hau-Ren
Lin Jui-Yen
Tsai Wei-Yü
Cheng Soo-Chen
References (42)
42 references, click to expand
  1. A new twist on RNA helicases: DExH/D box proteins as RNPases.
    Nat Struct Biol. 2001 Feb;8(2):113-6 PMID: 11175897
  2. 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
  3. Eviction and transplacement of mutant genes in yeast.
    Methods Enzymol. 1983;101:211-28 PMID: 6310325
  4. Yeast mRNA splicing in vitro.
    J Biol Chem. 1985 Nov 25;260(27):14780-92 PMID: 2997224
  5. Specific small nuclear RNAs are associated with yeast spliceosomes.
    Cell. 1986 Jun 20;45(6):869-77 PMID: 3518951
  6. Electrophoretic separation of complexes involved in the splicing of precursors to mRNAs.
    Cell. 1986 Sep 12;46(6):845-55 PMID: 2944598
  7. Electrophoresis of ribonucleoproteins reveals an ordered assembly pathway of yeast splicing complexes.
    Nature. 1986 Nov 27-Dec 3;324(6095):341-5 PMID: 3537805
  8. Interactions between small nuclear ribonucleoprotein particles in formation of spliceosomes.
    Cell. 1987 Jun 19;49(6):763-74 PMID: 2953438
  9. An ordered pathway of snRNP binding during mammalian pre-mRNA splicing complex assembly.
    EMBO J. 1987 Aug;6(8):2415-24 PMID: 2959470
  10. Spliceosome assembly in yeast.
    Genes Dev. 1987 Nov;1(9):1014-27 PMID: 2962902
  11. Spliceosome assembly involves the binding and release of U4 small nuclear ribonucleoprotein.
    Proc Natl Acad Sci U S A. 1988 Jan;85(2):411-5 PMID: 2963332
  12. The PRP4 (RNA4) protein of Saccharomyces cerevisiae is associated with the 5' portion of the U4 small nuclear RNA.
    Mol Cell Biol. 1990 Mar;10(3):1217-25 PMID: 2154681
  13. Requirement of the RNA helicase-like protein PRP22 for release of messenger RNA from spliceosomes.
    Nature. 1991 Feb 7;349(6309):487-93 PMID: 1992352
  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. Messenger RNA splicing in yeast: clues to why the spliceosome is a ribonucleoprotein.
    Science. 1991 Jul 12;253(5016):157-63 PMID: 1853200
  16. Association of U6 snRNA with the 5'-splice site region of pre-mRNA in the spliceosome.
    Genes Dev. 1992 Feb;6(2):244-54 PMID: 1310665
  17. U5 snRNA interacts with exon sequences at 5' and 3' splice sites.
    Cell. 1992 Feb 21;68(4):743-54 PMID: 1739979
  18. Interactions of small nuclear RNA's with precursor messenger RNA during in vitro splicing.
    Science. 1992 Sep 25;257(5078):1918-25 PMID: 1411506
  19. 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
  20. 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
  21. PRP19: a novel spliceosomal component.
    Mol Cell Biol. 1993 Mar;13(3):1876-82 PMID: 8441419
  22. The yeast PRP19 protein is not tightly associated with small nuclear RNAs, but appears to associate with the spliceosome after binding of U2 to the pre-mRNA and prior to formation of the functional spliceosome.
    Mol Cell Biol. 1993 Mar;13(3):1883-91 PMID: 7680101
  23. Yeast protein splicing factors involved in nuclear pre-mRNA splicing.
    Mol Biol Rep. 1993 Aug;18(2):99-103 PMID: 8232301
  24. 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
  25. Functional association of essential splicing factor(s) with PRP19 in a protein complex.
    EMBO J. 1994 May 15;13(10):2421-31 PMID: 8194532
  26. Split genes and RNA splicing.
    Cell. 1994 Jun 17;77(6):805-15 PMID: 7516265
  27. An evolutionarily conserved U5 snRNP-specific protein is a GTP-binding factor closely related to the ribosomal translocase EF-2.
    EMBO J. 1997 Jul 1;16(13):4092-106 PMID: 9233818
  28. Mechanical devices of the spliceosome: motors, clocks, springs, and things.
    Cell. 1998 Feb 6;92(3):315-26 PMID: 9476892
  29. 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
  30. Prp22, a DExH-box RNA helicase, plays two distinct roles in yeast pre-mRNA splicing.
    EMBO J. 1998 Apr 1;17(7):2086-94 PMID: 9524130
  31. The human U5-200kD DEXH-box protein unwinds U4/U6 RNA duplices in vitro.
    Proc Natl Acad Sci U S A. 1998 Apr 14;95(8):4188-92 PMID: 9539711
  32. The DEAH-box splicing factor Prp16 unwinds RNA duplexes in vitro.
    Curr Biol. 1998 Apr 9;8(8):441-51 PMID: 9550699
  33. The DEAH-box protein PRP22 is an ATPase that mediates ATP-dependent mRNA release from the spliceosome and unwinds RNA duplexes.
    EMBO J. 1998 May 15;17(10):2926-37 PMID: 9582286
  34. Myb-related Schizosaccharomyces pombe cdc5p is structurally and functionally conserved in eukaryotes.
    Mol Cell Biol. 1998 Jul;18(7):4097-108 PMID: 9632794
  35. An RNA switch at the 5' splice site requires ATP and the DEAD box protein Prp28p.
    Mol Cell. 1999 Jan;3(1):55-64 PMID: 10024879
  36. The identification and characterization of a novel splicing protein, Isy1p, of Saccharomyces cerevisiae.
    RNA. 1999 Mar;5(3):360-8 PMID: 10094305
  37. 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
  38. 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
  39. Yeast ortholog of the Drosophila crooked neck protein promotes spliceosome assembly through stable U4/U6.U5 snRNP addition.
    RNA. 1999 Aug;5(8):1042-54 PMID: 10445879
  40. Functional analyses of interacting factors involved in both pre-mRNA splicing and cell cycle progression in Saccharomyces cerevisiae.
    RNA. 2000 Nov;6(11):1565-72 PMID: 11105756
  41. Genetic and physical interactions between factors involved in both cell cycle progression and pre-mRNA splicing in Saccharomyces cerevisiae.
    Genetics. 2000 Dec;156(4):1503-17 PMID: 11102353
  42. Specific alterations of U1-C protein or U1 small nuclear RNA can eliminate the requirement of Prp28p, an essential DEAD box splicing factor.
    Mol Cell. 2001 Jan;7(1):227-32 PMID: 11172727
Article Info
Journal
Nucleic acids research
Abbr.
Nucleic Acids Res
ISSN
1362-4962
DOI
10.1093/nar/30.4.1029
Published
2002-02-15
Pages
1029-37
Language
English
Region
England
NLM ID
0411011
PMCID
PMC100336
Subset
IM
External Links
PubMed source DOI: 10.1093/nar/30.4.1029 PMC Full Text View journal details
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