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PMID: 1332064 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

The 5' splice site consensus RNA oligonucleotide induces assembly of U2/U4/U5/U6 small nuclear ribonucleoprotein complexes.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 89 ·No. 22 ·1992-11-15 ·Pages 10969-73

Hall KB, Konarska MM

Abstract

A short RNA oligonucleotide comprising the 5' splice site consensus sequence (5'SS RNA oligo) efficiently inhibits splicing of mRNA precursors in HeLa cell nuclear extracts. Addition of 5'SS RNA oligo inhibits early, but not late, steps in the splicing reaction, affecting the process of spliceosome assembly. In the presence of 5'SS RNA oligo a majority of U4/U5/U6 triple small nuclear ribonucleoprotein (snRNP) complex present in HeLa nuclear extracts associates with U2 snRNP to form a multi-snRNP complex, which could account for the observed inhibition of splicing by the oligo. This same set of snRNPs has been shown to assemble on pre-mRNAs during in vitro splicing to form splicing complex B. Removal of the 5' end of U1 snRNA, which is complementary to the 5' splice site, does not prevent association of snRNPs into U2/U4/U5/U6 complex in the presence of 5'SS RNA oligo. This suggests that interactions other than U1 snRNA.5'SS RNA oligo base pairing are used in recognition of the oligo sequence. 5'SS RNA oligo-induced assembly of the multi-snRNP complex may thus serve as a model to study the mechanism of 5' splice site recognition during splicing.

MeSH Terms
Base Sequence Cell Nucleus/metabolism HeLa Cells Humans Macromolecular Substances Molecular Sequence Data Oligoribonucleotides/pharmacology RNA Precursors/genetics,metabolism RNA Splicing/drug effects RNA, Small Nuclear/biosynthesis,genetics Ribonuclease H Ribonucleoproteins, Small Nuclear/biosynthesis,genetics
Chemicals
Macromolecular Substances Oligoribonucleotides RNA Precursors RNA, Small Nuclear Ribonucleoproteins, Small Nuclear Ribonuclease H
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Hall K B
Department of Biochemistry and Molecular Biophysics, Washington University School of Medicine, St. Louis, MO 63110.
Konarska M M
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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.89.22.10969
Published
1992-11-15
Pages
10969-73
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC50464
Subset
IM
External Links
PubMed source DOI: 10.1073/pnas.89.22.10969 PMC Full Text View journal details
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