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PMID: 9799242 Published · ppublish English Journal Article

The yeast U2A'/U2B complex is required for pre-spliceosome formation.

The EMBO journal ·Vol. 17 ·No. 21 ·1998-11-02 ·Pages 6348-58

Caspary F, Séraphin B

Abstract

Human U2 snRNP contains two specific proteins, U2A' and U2B", that interact with U2 snRNA stem-loop IV. In Saccharomyces cerevisiae, only the counterpart of human U2B", Yib9p, has been identified. Database searches revealed a gene potentially coding for a protein with striking similarities to human U2A', henceforth called LEA1 (looks exceptionally like U2A'). We demonstrate that Lea1p is a specific component of the yeast U2 snRNP. In addition, we show that Lea1p interacts directly with Yib9p. In vivo association of Lea1p with U2 snRNA requires Yib9p. Reciprocally, Yib9p binds to the U2 snRNA only in the presence of Lea1p in vivo, even though it has been previously shown to associate on its own with the U2 snRNA stem-loop IV in vitro. Strains lacking LEA1 and/or YIB9 grow slowly, are temperature sensitive and contain reduced levels of U2 snRNA. Pre-mRNA splicing is strongly impaired in these cells. In vitro studies demonstrate that spliceosome assembly is blocked prior to addition of U2 snRNP. This phenotype can be rescued partially, but specifically, by addition of the corresponding recombinant protein(s). This demonstrates a specific role for the yeast U2 snRNP specific proteins during formation of the pre-spliceosome.

MeSH Terms
Amino Acid Sequence Autoantigens Cell Division/genetics Databases as Topic Fungal Proteins/chemistry,genetics Genes, Fungal/genetics Molecular Sequence Data RNA Splicing/genetics RNA, Small Nuclear/genetics RNA-Binding Proteins/chemistry,genetics Ribonucleoprotein, U2 Small Nuclear/chemistry,genetics,metabolism Ribonucleoproteins, Small Nuclear Saccharomyces cerevisiae/genetics Saccharomyces cerevisiae Proteins Sequence Alignment Spliceosomes/genetics snRNP Core Proteins
Chemicals
Autoantigens Fungal Proteins LEA1 protein, S cerevisiae MSL1 protein, S cerevisiae RNA, Small Nuclear RNA-Binding Proteins Ribonucleoprotein, U2 Small Nuclear Ribonucleoproteins, Small Nuclear SNRPB2 protein, human Saccharomyces cerevisiae Proteins U2A' protein, human YIB9 protein, S cerevisiae snRNP Core Proteins
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Caspary F
EMBL, Meyerhofstrasse 1, D-69117 Heidelberg, Germany.
Séraphin B
References (59)
59 references, click to expand
  1. Amino acid sequence of the ribonuclease inhibitor from porcine liver reveals the presence of leucine-rich repeats.
    Biochemistry. 1988 Nov 15;27(23):8537-44 PMID: 3219361
  2. The prespliceosome components SAP 49 and SAP 145 interact in a complex implicated in tethering U2 snRNP to the branch site.
    Genes Dev. 1994 Aug 15;8(16):1974-83 PMID: 7958871
  3. A novel genetic system to detect protein-protein interactions.
    Nature. 1989 Jul 20;340(6230):245-6 PMID: 2547163
  4. Functional analysis of mutant Xenopus U2 snRNAs.
    Cell. 1989 Oct 6;59(1):159-69 PMID: 2790958
  5. Identification of functional U1 snRNA-pre-mRNA complexes committed to spliceosome assembly and splicing.
    Cell. 1989 Oct 20;59(2):349-58 PMID: 2529976
  6. U2 snRNA sequences that bind U2-specific proteins are dispensable for the function of U2 snRNP in splicing.
    Genes Dev. 1989 Dec;3(12A):1887-98 PMID: 2559872
  7. In vitro reconstitution of functional yeast U2 snRNPs.
    Genes Dev. 1989 Dec;3(12B):2124-36 PMID: 2560754
  8. Human U2 snRNA can function in pre-mRNA splicing in yeast.
    Nature. 1990 May 17;345(6272):270-3 PMID: 2185425
  9. Major determinants of the specificity of interaction between small nuclear ribonucleoproteins U1A and U2B'' and their cognate RNAs.
    Nature. 1990 Jun 7;345(6275):502-6 PMID: 2140872
  10. Structure of spliceosomal snRNPs and their role in pre-mRNA splicing.
    Biochim Biophys Acta. 1990 Nov 30;1087(3):265-92 PMID: 2147394
  11. Recognition of U1 and U2 small nuclear RNAs can be altered by a 5-amino-acid segment in the U2 small nuclear ribonucleoprotein particle (snRNP) B" protein and through interactions with U2 snRNP-A' protein.
    Mol Cell Biol. 1991 Apr;11(4):1829-39 PMID: 1826042
  12. The yeast branchpoint sequence is not required for the formation of a stable U1 snRNA-pre-mRNA complex and is recognized in the absence of U2 snRNA.
    EMBO J. 1991 May;10(5):1209-16 PMID: 1827069
  13. Cloning of a yeast U1 snRNP 70K protein homologue: functional conservation of an RNA-binding domain between humans and yeast.
    EMBO J. 1991 Sep;10(9):2627-34 PMID: 1714384
  14. Who's on first? The U1 snRNP-5' splice site interaction and splicing.
    Trends Biochem Sci. 1991 May;16(5):187-90 PMID: 1882420
  15. An ATP-independent complex commits pre-mRNA to the mammalian spliceosome assembly pathway.
    Genes Dev. 1991 Dec;5(12B):2534-46 PMID: 1836445
  16. Requirements for U2 snRNP addition to yeast pre-mRNA.
    Nucleic Acids Res. 1992 Aug 25;20(16):4237-45 PMID: 1387205
  17. Purification of splicing factor SF1, a heat-stable protein that functions in the assembly of a presplicing complex.
    Mol Cell Biol. 1992 Oct;12(10):4545-52 PMID: 1406644
  18. Small nuclear ribonucleoprotein (RNP) U2 contains numerous additional proteins and has a bipartite RNP structure under splicing conditions.
    Mol Cell Biol. 1993 Jan;13(1):307-19 PMID: 8380223
  19. The trans-spliceosomal U2 snRNP protein 40K of Trypanosoma brucei: cloning and analysis of functional domains reveals homology to a mammalian snRNP protein.
    EMBO J. 1993 Mar;12(3):1239-48 PMID: 8458336
  20. A simple and efficient method for direct gene deletion in Saccharomyces cerevisiae.
    Nucleic Acids Res. 1993 Jul 11;21(14):3329-30 PMID: 8341614
  21. Separation of splicing factor SF3 into two components and purification of SF3a activity.
    J Biol Chem. 1993 Aug 15;268(23):17640-6 PMID: 8349644
  22. Evidence that the 60-kDa protein of 17S U2 small nuclear ribonucleoprotein is immunologically and functionally related to the yeast PRP9 splicing factor and is required for the efficient formation of prespliceosomes.
    Proc Natl Acad Sci U S A. 1993 Sep 1;90(17):8229-33 PMID: 8367487
  23. Interaction of mammalian splicing factor SF3a with U2 snRNP and relation of its 60-kD subunit to yeast PRP9.
    Science. 1993 Oct 1;262(5130):102-5 PMID: 8211112
  24. Correspondence between a mammalian spliceosome component and an essential yeast splicing factor.
    Science. 1993 Oct 1;262(5130):105-8 PMID: 8211113
  25. Crystal structure at 1.92 A resolution of the RNA-binding domain of the U1A spliceosomal protein complexed with an RNA hairpin.
    Nature. 1994 Dec 1;372(6505):432-8 PMID: 7984237
  26. Sm and Sm-like proteins belong to a large family: identification of proteins of the U6 as well as the U1, U2, U4 and U5 snRNPs.
    EMBO J. 1995 May 1;14(9):2089-98 PMID: 7744014
  27. Nucleotide sequence and analysis of the centromeric region of yeast chromosome IX.
    Yeast. 1995 Jan;11(1):61-78 PMID: 7762303
  28. A method for performing precise alterations in the yeast genome using a recycable selectable marker.
    Nucleic Acids Res. 1995 Aug 11;23(15):3079-81 PMID: 7659536
  29. 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
  30. Evidence that sequence-independent binding of highly conserved U2 snRNP proteins upstream of the branch site is required for assembly of spliceosomal complex A.
    Genes Dev. 1996 Jan 15;10(2):233-43 PMID: 8566756
  31. Drosophila SNF/D25 combines the functions of the two snRNP proteins U1A and U2B' that are encoded separately in human, potato, and yeast.
    RNA. 1996 Jan;2(1):11-23 PMID: 8846293
  32. Identification and characterization of a yeast gene encoding the U2 small nuclear ribonucleoprotein particle B" protein.
    Mol Cell Biol. 1996 Jun;16(6):2787-95 PMID: 8649387
  33. The structure and function of proteins involved in mammalian pre-mRNA splicing.
    Annu Rev Biochem. 1996;65:367-409 PMID: 8811184
  34. The role of branchpoint-3' splice site spacing and interaction between intron terminal nucleotides in 3' splice site selection in Saccharomyces cerevisiae.
    EMBO J. 1997 Feb 17;16(4):779-92 PMID: 9049307
  35. Cross-intron bridging interactions in the yeast commitment complex are conserved in mammals.
    Cell. 1997 May 2;89(3):403-12 PMID: 9150140
  36. Protein functions in pre-mRNA splicing.
    Curr Opin Cell Biol. 1997 Jun;9(3):320-8 PMID: 9159080
  37. Toward a functional analysis of the yeast genome through exhaustive two-hybrid screens.
    Nat Genet. 1997 Jul;16(3):277-82 PMID: 9207794
  38. The CLUSTAL_X windows interface: flexible strategies for multiple sequence alignment aided by quality analysis tools.
    Nucleic Acids Res. 1997 Dec 15;25(24):4876-82 PMID: 9396791
  39. Conservation of structure and subunit interactions in yeast homologues of splicing factor 3b (SF3b) subunits.
    RNA. 1998 Jan;4(1):1-10 PMID: 9436903
  40. Mechanical devices of the spliceosome: motors, clocks, springs, and things.
    Cell. 1998 Feb 6;92(3):315-26 PMID: 9476892
  41. Expression of a beta-galactosidase gene containing the ribosomal protein 51 intron is sensitive to the rna2 mutation of yeast.
    Proc Natl Acad Sci U S A. 1983 Jul;80(14):4403-7 PMID: 6308621
  42. A comprehensive set of sequence analysis programs for the VAX.
    Nucleic Acids Res. 1984 Jan 11;12(1 Pt 1):387-95 PMID: 6546423
  43. mRNA splicing efficiency in yeast and the contribution of nonconserved sequences.
    Cell. 1985 May;41(1):119-26 PMID: 3888403
  44. Mutations in a yeast intron demonstrate the importance of specific conserved nucleotides for the two stages of nuclear mRNA splicing.
    Cell. 1986 Apr 11;45(1):81-93 PMID: 3513966
  45. An improved technique for the efficient construction of gene libraries by partial filling-in of cohesive ends.
    Gene. 1986;42(1):119-23 PMID: 3013727
  46. cDNA cloning of the human U1 snRNA-associated A protein: extensive homology between U1 and U2 snRNP-specific proteins.
    EMBO J. 1987 Dec 1;6(12):3841-8 PMID: 2962859
  47. 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
  48. Interactions within the yeast Sm core complex: from proteins to amino acids.
    Mol Cell Biol. 1998 Apr;18(4):1956-66 PMID: 9528767
  49. Phosphorylation of spliceosomal protein SAP 155 coupled with splicing catalysis.
    Genes Dev. 1998 May 15;12(10):1409-14 PMID: 9585501
  50. Crystal structure of the spliceosomal U2B"-U2A' protein complex bound to a fragment of U2 small nuclear RNA.
    Nature. 1998 Aug 13;394(6694):645-50 PMID: 9716128
  51. New constructs and strategies for efficient PCR-based gene manipulations in yeast.
    Yeast. 1998 Sep 15;14(12):1139-46 PMID: 9778799
  52. Interaction between PRP11 and SPP91 yeast splicing factors and characterization of a PRP9-PRP11-SPP91 complex.
    Science. 1993 Oct 1;262(5130):108-10 PMID: 8211114
  53. Crystal structure of porcine ribonuclease inhibitor, a protein with leucine-rich repeats.
    Nature. 1993 Dec 23-30;366(6457):751-6 PMID: 8264799
  54. Involvement of U6 snRNA in 5' splice site selection.
    Science. 1993 Dec 24;262(5142):2035-9 PMID: 8266100
  55. Parameters affecting lithium acetate-mediated transformation of Saccharomyces cerevisiae and development of a rapid and simplified procedure.
    Curr Genet. 1993 Nov;24(5):455-9 PMID: 8299163
  56. Conserved structures and diversity of functions of RNA-binding proteins.
    Science. 1994 Jul 29;265(5172):615-21 PMID: 8036511
  57. The POP1 gene encodes a protein component common to the RNase MRP and RNase P ribonucleoproteins.
    Genes Dev. 1994 Jun 15;8(12):1423-33 PMID: 7926742
  58. The yeast MUD2 protein: an interaction with PRP11 defines a bridge between commitment complexes and U2 snRNP addition.
    Genes Dev. 1994 Apr 1;8(7):843-54 PMID: 7926772
  59. Molecular cloning of the cDNA for the human U2 snRNA-specific A' protein.
    Nucleic Acids Res. 1989 Mar 11;17(5):1893-906 PMID: 2928112
Article Info
Journal
The EMBO journal
Abbr.
EMBO J
ISSN
0261-4189
DOI
10.1093/emboj/17.21.6348
Published
1998-11-02
Pages
6348-58
Language
English
Region
England
NLM ID
8208664
PMCID
PMC1170959
Subset
IM
External Links
PubMed source DOI: 10.1093/emboj/17.21.6348 PMC Full Text View journal details
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