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PMID: 11421357 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

3' splice site recognition in nematode trans-splicing involves enhancer-dependent recruitment of U2 snRNP.

RNA (New York, N.Y.) ·Vol. 7 ·No. 6 ·2001-06-00 ·Pages 785-92

Romfo CM, Maroney PA, Wu S, Nilsen TW

Abstract

Trans-splicing requires that 5' and 3' splice sites be independently recognized. Here, we have used mutational analyses and a sensitive nuclease protection assay to determine the mechanism of trans-3' splice site recognition in vitro. Efficient recognition of the 3' splice site is dependent upon both the sequence of the 3' splice site itself and enhancer elements located in the 3' exon. We show that the presence of three distinct classes of enhancers results in increased binding of U2 snRNP to the branchpoint region. Several lines of evidence strongly suggest that the increased binding of U2 snRNP is mediated by U2AF. These results expand the roles of enhancers in constitutive splicing and provide direct support for the recruitment model of enhancer function.

MeSH Terms
Animals Enhancer Elements, Genetic Exons Mutation Nematoda/genetics RNA Precursors/metabolism RNA Splicing RNA, Messenger/metabolism Ribonucleoprotein, U2 Small Nuclear/genetics,metabolism
Chemicals
RNA Precursors RNA, Messenger Ribonucleoprotein, U2 Small Nuclear
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Romfo C M
Center for RNA Molecular Biology, Department of Molecular Biology and Microbiology, Case Western Reserve University School of Medicine, Cleveland, Ohio 44106, USA.
Maroney P A
Wu S
Nilsen T W
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Article Info
Journal
RNA (New York, N.Y.)
Abbr.
RNA
ISSN
1355-8382
DOI
10.1017/s1355838201010263
Published
2001-06-00
Pages
785-92
Language
English
Region
United States
NLM ID
9509184
PMCID
PMC1370130
Subset
IM
Grants
NIA NIH HHS · AG0001015-15 · United States
NIAID NIH HHS · AI-28799 · United States
NIGMS NIH HHS · GM 31528 · United States
External Links
PubMed source DOI: 10.1017/s1355838201010263 PMC Full Text View journal details
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