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PMID: 1656228 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

Splice site choice in a complex transcription unit containing multiple inefficient polyadenylation signals.

Molecular and cellular biology ·Vol. 11 ·No. 10 ·1991-10-00 ·Pages 5291-300

Luo Y, Carmichael GG

Abstract

The relationship between polyadenylation and splicing was investigated in a model system consisting of two tandem but nonidentical polyomavirus late transcription units. This model system exploits the polyomavirus late transcription termination and polyadenylation signals, which are sufficiently weak to allow the production of many multigenome-length primary transcripts with repeating introns, exons, and poly(A) sites. This double-genome construct contains exons of two types, those bordered by 3' and 5' splice sites (L1 and L2) and those bordered by a 3' splice site and a poly(A) site (V1 and V2). The L1 and L2 exons are distinguishable from one another but retain identical flanking RNA processing signals, as is the case for the V1 and V2 exons. Analysis of cytoplasmic RNAs obtained from mouse cells transfected with this construct and its derivatives revealed the following. (i) V1 and V2 exons are often skipped during pre-mRNA processing, while L1 and L2 exons are not skipped. (ii) No messages contain internal, unused polyadenylation signals. (iii) Poly(A) site choice is not required for the selection of an upstream 3' splice site. (iv) When two tandem poly(A) sites are placed downstream of a 3' splice site, the first poly(A) site is chosen almost exclusively, even though transcription can proceed past both sites. (v) Placing a 3' splice site between these two tandem poly(A) sites allows the more distal site to be chosen. These and other available data are most consistent with a model in which terminal exons are produced by the coordinate selection and use of a 3' splice site with the nearest available downstream poly(A) site.

MeSH Terms
Animals Base Sequence Cell Line Cloning, Molecular Exons/genetics Mice Molecular Sequence Data Poly A/genetics Polymerase Chain Reaction Polyomavirus/genetics RNA Precursors/metabolism RNA Probes/genetics RNA Splicing/genetics RNA, Messenger/metabolism Repetitive Sequences, Nucleic Acid/genetics
Chemicals
RNA Precursors RNA Probes RNA, Messenger Poly A
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Luo Y
Department of Microbiology, University of Connecticut Health Center, Farmington 06030.
Carmichael G G
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56 references, click to expand
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Article Info
Journal
Molecular and cellular biology
Abbr.
Mol Cell Biol
ISSN
0270-7306
DOI
10.1128/mcb.11.10.5291-5300.1991
Published
1991-10-00
Pages
5291-300
Language
English
Region
United States
NLM ID
8109087
PMCID
PMC361585
Subset
IM
Grants
NCI NIH HHS · R01 CA045382 · United States
NCI NIH HHS · CA45382 · United States
External Links
PubMed source DOI: 10.1128/mcb.11.10.5291-5300.1991 PMC Full Text View journal details
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