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PMID: 8649407 Published · ppublish English Journal Article

cis-acting sequences located downstream of the human immunodeficiency virus type 1 promoter affect its chromatin structure and transcriptional activity.

Molecular and cellular biology ·Vol. 16 ·No. 6 ·1996-06-00 ·Pages 2958-66

el Kharroubi A, Martin MA

Abstract

We have examined the roles of AP-1, AP-3-like, DBF1, and Sp1 binding sites, which are located downstream of the human immunodeficiency virus type 1 (HIV-1) promoter, in regulating basal transcriptional activity directed by the integrated viral long terminal repeat (LTR). Point mutations affecting all four of these elements functionally inactivated the HIV-1 LTR when it was constrained in a chromatin configuration. Analyses of the chromatin structures of the transcriptionally active wild-type and inactive mutated HIV-1 promoters revealed several differences. In the active promoter, the 3' half of the U3 region, including the basal promoter, the enhancer, and the putative upstream regulatory sequences are situated within a nuclease-hypersensitive region. However, the far upstream U3 region appears to be packaged into a nuclease-resistant nucleosomal structure, whereas the R, U5, and gag leader sequences are associated with a region of altered chromatin that is sensitive to restriction endonucleases. In the inactive template, only the basal promoter and enhancer element remain sensitive to nucleases, and the adjacent upstream and downstream regions are incorporated into nuclease-resistant nucleosomal structures. Taken together, these results indicate that the chromatin structure of the integrated HIV-1 LTR plays a critical role in modulating basal transcriptional activity.

MeSH Terms
Base Sequence Binding Sites/genetics Cell Line Chloramphenicol O-Acetyltransferase/genetics Chromatin/genetics,metabolism Chromosome Mapping DNA Primers/genetics DNA, Viral/genetics,metabolism Genes, Reporter Genes, Viral Genes, gag HIV Long Terminal Repeat HIV-1/genetics HeLa Cells Humans Molecular Sequence Data Point Mutation Promoter Regions, Genetic Sequence Deletion Transcription Factors/metabolism Transcription, Genetic
Chemicals
Chromatin DNA Primers DNA, Viral Transcription Factors Chloramphenicol O-Acetyltransferase
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
el Kharroubi A
Laboratory of Molecular Microbiology, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, Maryland 20892, USA.
Martin M A
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Article Info
Journal
Molecular and cellular biology
Abbr.
Mol Cell Biol
ISSN
0270-7306
DOI
10.1128/MCB.16.6.2958
Published
1996-06-00
Pages
2958-66
Language
English
Region
United States
NLM ID
8109087
PMCID
PMC231290
Subset
IM
External Links
PubMed source DOI: 10.1128/MCB.16.6.2958 PMC Full Text View journal details
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