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PMID: 8035806 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Sequences homologous to 5' splice sites are required for the inhibitory activity of papillomavirus late 3' untranslated regions.

Molecular and cellular biology ·Vol. 14 ·No. 8 ·1994-08-00 ·Pages 5278-89

Furth PA, Choe WT, Rex JH, Byrne JC, Baker CC

Abstract

Expression of bovine papillomavirus type 1 (BPV-1) late genes is limited to terminally differentiated keratinocytes in an infected epithelium. We have previously shown that although the BPV-1 late polyadenylation site is functional in nonpermissive cells, a 53-nucleotide (nt) fragment of the late 3' untranslated region acts posttranscriptionally to reduce polyadenylated cytoplasmic RNA levels. This 53-nt fragment does not appear to function by destabilizing polyadenylated cytoplasmic RNA (P. A. Furth and C. C. Baker, J. Virol. 65:5806-5812, 1991). In this study, we used site-directed mutagenesis and deletion analysis to demonstrate that the sequence AAG/GUAAGU, which is identical to the consensus 5' splice site sequence, was both necessary and sufficient for the inhibitory activity of the 53-nt fragment. Furthermore, base pairing between the 5' end of the U1 small nuclear RNA and this 5' splice site-like sequence was shown to be required for the inhibitory activity in vivo. We have also further mapped the human papillomavirus type 16 late 3' inhibitory element (I. M. Kennedy, J. K. Haddow, and J. B. Clements, J. Virol. 65:2093-2097, 1991) to a 51-nt region containing four overlapping sequence motifs with partial homology to 5' splice sites. Mutation of each of these motifs demonstrated that only one of these motifs is required for the inhibitory activity. However, the presence of the other motifs may contribute to the full inhibitory activity of the element. No BPV-1 or human papillomavirus type 16 mRNAs which are spliced by using the potential 5' splice sites present in the viral late 3' untranslated regions have been identified. This suggests that the primary function of these 5' splice site-like sequences is the inhibition of late gene expression. The most likely mechanism of action of these elements is reduction of polyadenylation efficiency, perhaps through interference with 3'-terminal exon definition.

MeSH Terms
Base Sequence Bovine papillomavirus 1/genetics Consensus Sequence Gene Expression Regulation, Viral Hydrogen Bonding Molecular Sequence Data Mutagenesis, Site-Directed Papillomaviridae/genetics Poly A/metabolism RNA Processing, Post-Transcriptional RNA Splicing RNA, Messenger/genetics RNA, Small Nuclear/genetics RNA, Viral/genetics Regulatory Sequences, Nucleic Acid Structure-Activity Relationship
Chemicals
RNA, Messenger RNA, Small Nuclear RNA, Viral Poly A
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Furth P A
Laboratory of Tumor Virus Biology, National Cancer Institute, Bethesda, Maryland 20892.
Choe W T
Rex J H
Byrne J C
Baker C C
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Article Info
Journal
Molecular and cellular biology
Abbr.
Mol Cell Biol
ISSN
0270-7306
DOI
10.1128/mcb.14.8.5278-5289.1994
Published
1994-08-00
Pages
5278-89
Language
English
Region
United States
NLM ID
8109087
PMCID
PMC359047
Subset
IM
External Links
PubMed source DOI: 10.1128/mcb.14.8.5278-5289.1994 PMC Full Text View journal details
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