Home LiteratureArticle Details
PMID: 2963332 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Spliceosome assembly involves the binding and release of U4 small nuclear ribonucleoprotein.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 85 ·No. 2 ·1988-01-00 ·Pages 411-5

Lamond AI, Konarska MM, Grabowski PJ, Sharp PA

Abstract

Splicing complexes that form a rabbit beta-globin precursor mRNA (pre-mRNA) have been analyzed for their small nuclear RNA (snRNA) content by both affinity chromatography and specific probe hybridization of replicas of native electrophoretic gels. A pathway of spliceosome assembly was deduced that has at least three stages. (i) U2 small nuclear ribonucleoprotein (snRNP) alone binds to sequences of mRNA upstream of the 3' splice site. (ii) U4, U5, and U6 snRNPs bind, apparently simultaneously. (iii) U4 snRNP is released to generate a spliceosome that contains U2, U5, and U6 snRNPs together with the RNA intermediates in splicing. U1 snRNP was not detected in association with any of these complexes. A parallel analysis of the spliceosome found with an adenovirus precursor mRNA substrate yielded an identical snRNP composition with one additional, unidentified RNA species, called X. This latter RNA species was not detected in the spliceosome bound to the beta-globin substrate.

MeSH Terms
Cell Nucleus Globins/genetics HeLa Cells/metabolism Humans Molecular Weight RNA Precursors/genetics RNA Splicing RNA, Catalytic RNA, Ribosomal/genetics Ribonucleoproteins/genetics,isolation & purification,metabolism Ribonucleoproteins, Small Nuclear Transcription, Genetic
Chemicals
RNA Precursors RNA, Catalytic RNA, Ribosomal Ribonucleoproteins Ribonucleoproteins, Small Nuclear Globins
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Lamond A I
Center for Cancer Research, Massachusetts Institute of Technology, Cambridge 02139.
Konarska M M
Grabowski P J
Sharp P A
References (27)
27 references, click to expand
  1. Accurate transcription initiation by RNA polymerase II in a soluble extract from isolated mammalian nuclei.
    Nucleic Acids Res. 1983 Mar 11;11(5):1475-89 PMID: 6828386
  2. Spliceosome assembly in yeast.
    Genes Dev. 1987 Nov;1(9):1014-27 PMID: 2962902
  3. Evidence for the existence of snRNAs U4 and U6 in a single ribonucleoprotein complex and for their association by intermolecular base pairing.
    EMBO J. 1984 Jun;3(6):1357-63 PMID: 6204860
  4. The 5' terminus of the RNA moiety of U1 small nuclear ribonucleoprotein particles is required for the splicing of messenger RNA precursors.
    Cell. 1984 Aug;38(1):299-307 PMID: 6235919
  5. 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
  6. Molecular consequences of specific intron mutations on yeast mRNA splicing in vivo and in vitro.
    Cell. 1985 Aug;42(1):335-44 PMID: 3893746
  7. A multicomponent complex is involved in the splicing of messenger RNA precursors.
    Cell. 1985 Aug;42(1):345-53 PMID: 3160482
  8. Stepwise assembly of a pre-mRNA splicing complex requires U-snRNPs and specific intron sequences.
    Cell. 1985 Aug;42(1):355-67 PMID: 3160483
  9. 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
  10. U2 as well as U1 small nuclear ribonucleoproteins are involved in premessenger RNA splicing.
    Cell. 1985 Oct;42(3):737-50 PMID: 2996775
  11. 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
  12. Specific small nuclear RNAs are associated with yeast spliceosomes.
    Cell. 1986 Jun 20;45(6):869-77 PMID: 3518951
  13. U1, U2, and U4/U6 small nuclear ribonucleoproteins are required for in vitro splicing but not polyadenylation.
    Cell. 1986 Aug 29;46(5):691-6 PMID: 2427201
  14. Pre-mRNA splicing in vitro requires intact U4/U6 small nuclear ribonucleoprotein.
    Cell. 1986 Aug 29;46(5):697-704 PMID: 2427202
  15. Mutations in conserved intron sequences affect multiple steps in the yeast splicing pathway, particularly assembly of the spliceosome.
    EMBO J. 1986 Jul;5(7):1683-95 PMID: 3017708
  16. Affinity chromatography of splicing complexes: U2, U5, and U4 + U6 small nuclear ribonucleoprotein particles in the spliceosome.
    Science. 1986 Sep 19;233(4770):1294-9 PMID: 3638792
  17. A compensatory base change in U1 snRNA suppresses a 5' splice site mutation.
    Cell. 1986 Sep 12;46(6):827-35 PMID: 3757028
  18. Electrophoretic separation of complexes involved in the splicing of precursors to mRNAs.
    Cell. 1986 Sep 12;46(6):845-55 PMID: 2944598
  19. Sequence requirements for splicing of higher eukaryotic nuclear pre-mRNA.
    Cell. 1986 Nov 21;47(4):555-65 PMID: 3779836
  20. Ribonucleoprotein complex formation during pre-mRNA splicing in vitro.
    Mol Cell Biol. 1986 Jul;6(7):2582-92 PMID: 2946939
  21. Electrophoresis of ribonucleoproteins reveals an ordered assembly pathway of yeast splicing complexes.
    Nature. 1986 Nov 27-Dec 3;324(6095):341-5 PMID: 3537805
  22. Interactions between small nuclear ribonucleoprotein particles in formation of spliceosomes.
    Cell. 1987 Jun 19;49(6):763-74 PMID: 2953438
  23. 5' cleavage site in eukaryotic pre-mRNA splicing is determined by the overall 5' splice region, not by the conserved 5' GU.
    Cell. 1987 Jul 17;50(2):237-46 PMID: 3647844
  24. An essential snRNA from S. cerevisiae has properties predicted for U4, including interaction with a U6-like snRNA.
    Cell. 1987 Aug 14;50(4):585-92 PMID: 2440583
  25. An ordered pathway of snRNP binding during mammalian pre-mRNA splicing complex assembly.
    EMBO J. 1987 Aug;6(8):2415-24 PMID: 2959470
  26. A mutational analysis of spliceosome assembly: evidence for splice site collaboration during spliceosome formation.
    Genes Dev. 1987 Aug;1(6):532-43 PMID: 2824284
  27. U4 and U6 RNAs coexist in a single small nuclear ribonucleoprotein particle.
    Nucleic Acids Res. 1984 Apr 11;12(7):3283-93 PMID: 6201826
Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
DOI
10.1073/pnas.85.2.411
Published
1988-01-00
Pages
411-5
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC279558
Subset
IM
Grants
NCI NIH HHS · CA14051 · United States
NIGMS NIH HHS · GM34277 · United States
External Links
PubMed source DOI: 10.1073/pnas.85.2.411 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: [email protected]

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