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PMID: 8479905 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

The biochemical defects of prp4-1 and prp6-1 yeast splicing mutants reveal that the PRP6 protein is required for the accumulation of the [U4/U6.U5] tri-snRNP.

Nucleic acids research ·Vol. 21 ·No. 7 ·1993-04-11 ·Pages 1555-62

Galisson F, Legrain P

Abstract

We have raised specific antibodies against the PRP6 protein and shown that the U4, U5 and U6 snRNAs are co-precipitated with this protein. Using splicing extracts prepared from in vivo heat-inactivated cells, we have characterized the prp4-1 and prp6-1 biochemical defects. In inactivated prp4-1 cell extracts, the U6 snRNA content as well as the U6, U4/U6 snRNPs and the [U4/U6.U5] tri-snRNP particles amounts are severely reduced. In inactivated prp6-1 cell extracts, the PRP6 mutant protein is barely detectable. Glycerol gradient analyses indicate that, in these extracts, the [U4/U6.U5] tri-snRNPs are present in very low amounts, but U4/U6 snRNP particles are normally represented. These results establish that the PRP6 protein is required for the accumulation of the [U4/U6.U5] tri-snRNP. We found no evidence for the presence of the PRP6 protein in the U4/U6 particle.

Related Genes
prp4-1 prp6-1
MeSH Terms
Centrifugation, Density Gradient Fungal Proteins Glycerol Mutation Precipitin Tests RNA Splicing/genetics Ribonucleoprotein, U4-U6 Small Nuclear/metabolism Ribonucleoprotein, U5 Small Nuclear/metabolism Saccharomyces cerevisiae Saccharomyces cerevisiae Proteins Spliceosomes/metabolism
Chemicals
Fungal Proteins PRP6 protein, S cerevisiae Ribonucleoprotein, U4-U6 Small Nuclear Ribonucleoprotein, U5 Small Nuclear Saccharomyces cerevisiae Proteins Glycerol
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Galisson F
Département de Biologie Moléculaire, Institut Pasteur, Paris, France.
Legrain P
References (49)
49 references, click to expand
  1. Specific small nuclear RNAs are associated with yeast spliceosomes.
    Cell. 1986 Jun 20;45(6):869-77 PMID: 3518951
  2. A yeast splicing factor is localized in discrete subnuclear domains.
    EMBO J. 1992 Oct;11(10):3731-6 PMID: 1396568
  3. 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
  4. Electrophoresis of ribonucleoproteins reveals an ordered assembly pathway of yeast splicing complexes.
    Nature. 1986 Nov 27-Dec 3;324(6095):341-5 PMID: 3537805
  5. 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
  6. Interactions between small nuclear ribonucleoprotein particles in formation of spliceosomes.
    Cell. 1987 Jun 19;49(6):763-74 PMID: 2953438
  7. An ordered pathway of snRNP binding during mammalian pre-mRNA splicing complex assembly.
    EMBO J. 1987 Aug;6(8):2415-24 PMID: 2959470
  8. A trans-acting suppressor restores splicing of a yeast intron with a branch point mutation.
    Genes Dev. 1987 Jul;1(5):445-55 PMID: 2890553
  9. Vectors for selective expression of cloned DNAs by T7 RNA polymerase.
    Gene. 1987;56(1):125-35 PMID: 3315856
  10. Identification of a yeast snRNP protein and detection of snRNP-snRNP interactions.
    Cell. 1987 Dec 24;51(6):1019-26 PMID: 2961458
  11. Spliceosome assembly in yeast.
    Genes Dev. 1987 Nov;1(9):1014-27 PMID: 2962902
  12. RNA11 protein is associated with the yeast spliceosome and is localized in the periphery of the cell nucleus.
    Mol Cell Biol. 1988 Jun;8(6):2379-93 PMID: 3043176
  13. 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
  14. Early commitment of yeast pre-mRNA to the spliceosome pathway.
    Mol Cell Biol. 1988 Sep;8(9):3755-60 PMID: 3065622
  15. 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
  16. PRP4: a protein of the yeast U4/U6 small nuclear ribonucleoprotein particle.
    Mol Cell Biol. 1989 Sep;9(9):3710-9 PMID: 2528687
  17. Isolation and characterization of pre-mRNA splicing mutants of Saccharomyces cerevisiae.
    Genes Dev. 1989 Aug;3(8):1206-16 PMID: 2676722
  18. Identification of functional U1 snRNA-pre-mRNA complexes committed to spliceosome assembly and splicing.
    Cell. 1989 Oct 20;59(2):349-58 PMID: 2529976
  19. The mammalian analogue of the yeast PRP8 splicing protein is present in the U4/5/6 small nuclear ribonucleoprotein particle and the spliceosome.
    Proc Natl Acad Sci U S A. 1989 Nov;86(22):8742-6 PMID: 2479028
  20. Conservation between yeast and man of a protein associated with U5 small nuclear ribonucleoprotein.
    Nature. 1989 Dec 14;342(6251):819-21 PMID: 2532307
  21. 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
  22. PRP5: a helicase-like protein required for mRNA splicing in yeast.
    Proc Natl Acad Sci U S A. 1990 Jun;87(11):4236-40 PMID: 2349233
  23. The molecular characterization of PRP6 and PRP9 yeast genes reveals a new cysteine/histidine motif common to several splicing factors.
    EMBO J. 1990 Sep;9(9):2775-81 PMID: 2118103
  24. Targeted snRNP depletion reveals an additional role for mammalian U1 snRNP in spliceosome assembly.
    Cell. 1990 Oct 19;63(2):293-302 PMID: 2170025
  25. 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
  26. The yeast PRP6 gene encodes a U4/U6 small nuclear ribonucleoprotein particle (snRNP) protein, and the PRP9 gene encodes a protein required for U2 snRNP binding.
    Mol Cell Biol. 1990 Dec;10(12):6417-25 PMID: 2147224
  27. Structure of spliceosomal snRNPs and their role in pre-mRNA splicing.
    Biochim Biophys Acta. 1990 Nov 30;1087(3):265-92 PMID: 2147394
  28. Interactions of PRP2 protein with pre-mRNA splicing complexes in Saccharomyces cerevisiae.
    Nucleic Acids Res. 1990 Nov 25;18(22):6559-64 PMID: 2251118
  29. Identification of ten genes that control ribosome formation in yeast.
    Mol Gen Genet. 1970;109(1):42-56 PMID: 5488085
  30. Transformation of intact yeast cells treated with alkali cations.
    J Bacteriol. 1983 Jan;153(1):163-8 PMID: 6336730
  31. Genomic sequencing.
    Proc Natl Acad Sci U S A. 1984 Apr;81(7):1991-5 PMID: 6326095
  32. mRNA splicing efficiency in yeast and the contribution of nonconserved sequences.
    Cell. 1985 May;41(1):119-26 PMID: 3888403
  33. Molecular consequences of specific intron mutations on yeast mRNA splicing in vivo and in vitro.
    Cell. 1985 Aug;42(1):335-44 PMID: 3893746
  34. Multiple roles for U6 snRNA in the splicing pathway.
    Genes Dev. 1990 Dec;4(12B):2264-77 PMID: 2149118
  35. 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
  36. Cloning of the two essential yeast genes, PRP6 and PRP9, and their rapid mapping, disruption and partial sequencing using a linker insertion strategy.
    Mol Gen Genet. 1991 Feb;225(2):199-202 PMID: 1848649
  37. A cold-sensitive mRNA splicing mutant is a member of the RNA helicase gene family.
    Genes Dev. 1991 Apr;5(4):629-41 PMID: 2010088
  38. 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
  39. Pre-mRNA splicing in yeast.
    Trends Genet. 1991 Mar;7(3):79-85 PMID: 2031287
  40. Messenger RNA splicing in yeast: clues to why the spliceosome is a ribonucleoprotein.
    Science. 1991 Jul 12;253(5016):157-63 PMID: 1853200
  41. 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
  42. Immunoaffinity purification of a [U4/U6.U5] tri-snRNP from human cells.
    Genes Dev. 1991 Aug;5(8):1439-52 PMID: 1831175
  43. U4 small nuclear RNA dissociates from a yeast spliceosome and does not participate in the subsequent splicing reaction.
    Mol Cell Biol. 1991 Nov;11(11):5571-7 PMID: 1833635
  44. An ATP-independent complex commits pre-mRNA to the mammalian spliceosome assembly pathway.
    Genes Dev. 1991 Dec;5(12B):2534-46 PMID: 1836445
  45. A splicing factor that is inactivated during in vivo heat shock is functionally equivalent to the [U4/U6.U5] triple snRNP-specific proteins.
    Genes Dev. 1992 Apr;6(4):631-41 PMID: 1532785
  46. Biochemical mechanisms of constitutive and regulated pre-mRNA splicing.
    Annu Rev Cell Biol. 1991;7:559-99 PMID: 1839712
  47. PRP38 encodes a yeast protein required for pre-mRNA splicing and maintenance of stable U6 small nuclear RNA levels.
    Mol Cell Biol. 1992 Sep;12(9):3939-47 PMID: 1508195
  48. Roles of PRP8 protein in the assembly of splicing complexes.
    EMBO J. 1992 Oct;11(10):3721-9 PMID: 1396567
  49. The yeast RNA gene products are essential for mRNA splicing in vitro.
    Cell. 1986 Dec 26;47(6):953-63 PMID: 3536128
Article Info
Journal
Nucleic acids research
Abbr.
Nucleic Acids Res
ISSN
0305-1048
DOI
10.1093/nar/21.7.1555
Published
1993-04-11
Pages
1555-62
Language
English
Region
England
NLM ID
0411011
PMCID
PMC309362
Subset
IM
External Links
PubMed source DOI: 10.1093/nar/21.7.1555 PMC Full Text View journal details
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