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PMID: 3443104 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S. Research Support, U.S. Gov't, P.H.S.

Analysis of mRNA 3' end formation by modification interference: the only modifications which prevent processing lie in AAUAAA and the poly(A) site.

The EMBO journal ·Vol. 6 ·No. 13 ·1987-12-20 ·Pages 4177-84

Conway L, Wickens M

Abstract

A modification interference method is described in which chemically modified transcripts are used to identify bases required for any reaction for which synthetic RNA is a substrate. This technique provides information analogous to that obtained from the analysis of a complete set of point mutants. Using SV40 late pre-mRNAs, we determine that modification of any base in the AAUAAA sequence prevents cleavage, polyadenylation and formation of pre-cleavage complexes in vitro. Modification of the A to which poly(A) is added prevents polyadenylation, but does not interfere with formation of the pre-cleavage complex. No single modification downstream of the poly(A) site significantly affects cleavage efficiency. Since the region downstream of the poly(A) site is required for cleavage and complex formation (Conway and Wickens, 1985; Zarkower and Wickens, 1987b), we infer that the critical features of this downstream region are either diffuse or redundant.

MeSH Terms
Base Sequence Cell Nucleus/metabolism HeLa Cells/metabolism Humans Poly A RNA, Messenger/genetics Templates, Genetic Transcription, Genetic
Chemicals
RNA, Messenger Poly A
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Conway L
Department of Biochemistry, College of Agriculture and Life Sciences, University of Wisconsin, Madison 53706.
Wickens M
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35 references, click to expand
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Article Info
Journal
The EMBO journal
Abbr.
EMBO J
ISSN
0261-4189
Published
1987-12-20
Pages
4177-84
Language
English
Region
England
NLM ID
8208664
PMCID
PMC553901
Subset
IM
Grants
NIGMS NIH HHS · GM31892 · United States
External Links
PubMed source PMC Full Text View journal details
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