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PMID: 9469840 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Transcriptional interference perturbs the binding of Sp1 to the HIV-1 promoter.

Nucleic acids research ·Vol. 26 ·No. 5 ·1998-03-01 ·Pages 1294-301

Greger IH, Demarchi F, Giacca M, Proudfoot NJ

Abstract

Transcriptional interference between adjacent genes has been demonstrated in a variety of biological systems. To study this process in RNA polymerase II (pol II) transcribed genes we have analysed the effect of transcription on tandem HIV-1 promoters integrated into the genome of HeLa cells. We show that transcriptional activation at the upstream promoter reduces transcription from the downstream promoter, as compared with basal transcription conditions (in the absence of an activator). Furthermore, insertion of a strong transcriptional termination element between the two promoters alleviates this transcriptional interference, resulting in elevated levels of transcription from the downstream promoter. Actual protein interactions with the downstream (occluded) promoter were analysed by in vivo footprinting. Binding of Sp1 transcription factors to the occluded promoter was reduced, when compared with the footprint pattern of the promoter protected by the terminator. This suggests that promoter occlusion is due to disruption of certain transcription factors and that it can be blocked by an intervening transcriptional terminator. Chromatin mapping with DNase I indicates that a factor binds to the termination element under both basal and induced conditions.

MeSH Terms
Binding Sites/genetics Genes, tat Genome, Human HIV Long Terminal Repeat HIV-1/genetics HeLa Cells Humans Models, Genetic Plasmids/genetics Promoter Regions, Genetic Sp1 Transcription Factor/metabolism Transcription, Genetic Transcriptional Activation Transfection
Chemicals
Sp1 Transcription Factor
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Greger I H
Sir William Dunn School of Pathology, University of Oxford, South Parks Road, Oxford OX1 3RE, UK.
Demarchi F
Giacca M
Proudfoot N J
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Article Info
Journal
Nucleic acids research
Abbr.
Nucleic Acids Res
ISSN
0305-1048
DOI
10.1093/nar/26.5.1294
Published
1998-03-01
Pages
1294-301
Language
English
Region
England
NLM ID
0411011
PMCID
PMC147389
Subset
IM
Grants
Wellcome Trust · United Kingdom
External Links
PubMed source DOI: 10.1093/nar/26.5.1294 PMC Full Text View journal details
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