Cited by 125RNA Research and SplicingHIV Research and TreatmentRNA and protein synthesis mechanisms

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Poly(A) site selection in the HIV-1 provirus: inhibition of promoter-proximal polyadenylation by the downstream major splice donor site

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Ashe, M P, Griffin, P, James, W, Proudfoot, N J·1995·Genes & Development

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Abstract

In common with all retroviruses, the human immunodeficiency virus type 1 (HIV-1) contains duplicated long terminal repeat (LTR) sequences flanking the proviral genome. These LTRs contain identical poly(A) signals, which are both transcribed into RNA. Therefore, to allow efficient viral expression, a mechanism must exist to either restrict promoter-proximal poly(A) site use or enhance the activity of the promoter-distal poly(A) site. We have examined the use of both poly(A) sites using proviral clones. Mutation of the previously defined upstream activatory sequences of the 3' LTR poly(A) site decreases the efficiency of polyadenylation when placed in competition with an efficient downstream processing signal. However, in the absence of competition, these mutations have no effect on HIV-1 polyadenylation. In addition, the 5' LTR poly(A) site is inactive, whereas a heterologous poly(A) site positioned in its place is utilized efficiently. Furthermore, transcription initiating from the 3' LTR promoter utilizes the 3' LTR poly(A) signal efficiently. Therefore, the main determinant of the differential poly(A) site use appears to be neither proximity to a promoter element in the 5' LTR nor the presence of upstream activating sequences at the 3' LTR. Instead, we show that the major splice donor site that is immediately downstream of the 5' LTR inhibits cleavage and polyadenylation at the promoter-proximal site. The fact that this poly(A) site is active in a proviral clone when the major splice donor site is mutated suggests that the selective use of poly(A) signals in HIV-1 is mediated by a direct inhibition of the HIV-1 poly(A) site by downstream splicing events or factors involved in splicing.

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Authors: Ashe, M. P., Griffin, P., James, W., & Proudfoot, N. J.
Affiliations: University of Oxford (GB)
ORCID: Ashe, M P
Griffin, P
James, W
Proudfoot, N J
Venue: Genes & Development
Publisher: Cold Spring Harbor Laboratory
Published: 1995-12-01
Volume: 9
Issue: 23
Pages: 3008-3025
ISSN: 0890-9369 (ISSN-L)
1549-5477
DOI: 10.1101/gad.9.23.3008
OpenAlex: W2108413560
Cited by: 125
References: 62
Citations by year: 2026: 1 · 2025: 4 · 2024: 4 · 2023: 5 · 2022: 4 · 2021: 2 · 2019: 3 · 2018: 5 · 2017: 2 · 2016: 5 · 2015: 2 · 2014: 5 · 2013: 5 · 2012: 4
Type: posted content
Language: en
Open access: bronze
doi.org
Locations: Genes & Development (publishedVersion)
PubMed (publishedVersion)
Research Explorer (The University of Manchester) (submittedVersion)
Funders: Wellcome Trust
Medical Research Council
Topics: Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › RNA Research and Splicing
Life Sciences › Immunology and Microbiology › Virology › HIV Research and Treatment
Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › RNA and protein synthesis mechanisms
Keywords: HIV-1, polyadenylation site selection, long terminal repeat, upstream activator sequence, 5' LTR, 3' LTR, retroviral transcription, polyadenylation signal
MeSH terms: Base Sequence, Binding, Competitive, Gene Expression Regulation, Humans, Molecular Sequence Data, Mutation, Poly A, Promoter Regions, Genetic, Proviruses, RNA Splicing, RNA, Messenger, Transcription, Genetic, HIV-1, HIV Long Terminal Repeat

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Poly(A) site selection in the HIV-1 provirus: inhibition of promoter-proximal polyadenylation by the downstream major splice donor site

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