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PMID: 2545899 Published · ppublish English Journal Article

Functional characterization of a complex protein-DNA-binding domain located within the human immunodeficiency virus type 1 long terminal repeat leader region.

Journal of virology ·Vol. 63 ·No. 8 ·1989-08-00 ·Pages 3213-9

Malim MH, Fenrick R, Ballard DW, Hauber J, Böhnlein E, Cullen BR

Abstract

Transcriptional trans activation of the human immunodeficiency virus type 1 (HIV-1) long terminal repeat (LTR) by the viral tat trans activator is mediated by an LTR-specific sequence located immediately 3' to the start of transcription initiation. We have used a range of molecular techniques to examine DNA-protein interactions that occur in the vicinity of this cis-acting sequence. Our results demonstrate the existence of a sequence-specific DNA-protein interaction involving the HIV-1 leader DNA and map this binding event to between -2 and +21 base pairs relative to the HIV-1 LTR transcription start site. Evidence suggesting that this interaction involves three distinct protein-DNA contact sites extending along one side of the DNA helix is presented. Mutation of these sites was found to ablate protein-DNA binding yet was observed to have no effect on either the basal or tat trans-activated level of HIV-1 LTR-specific gene expression. We therefore conclude that this DNA-protein interaction has a function distinct from the regulation of HIV-1 LTR-specific gene expression.

MeSH Terms
Base Sequence Cell Line DNA Probes DNA, Viral/metabolism DNA-Binding Proteins/genetics,metabolism Gene Expression Regulation Gene Products, tat HIV-1/genetics HeLa Cells Humans Methylation Molecular Sequence Data Mutation Protein Binding Protein Sorting Signals/genetics Repetitive Sequences, Nucleic Acid Transcription Factors/genetics Transcription, Genetic Transfection Viral Proteins/genetics,metabolism tat Gene Products, Human Immunodeficiency Virus
Chemicals
DNA Probes DNA, Viral DNA-Binding Proteins Gene Products, tat Protein Sorting Signals Transcription Factors Viral Proteins tat Gene Products, Human Immunodeficiency Virus
Authors & Affiliations
6 authors, click to expand affiliations / ORCID
Malim M H
Howard Hughes Medical Institute, Duke University Medical Center, Durham, North Carolina 27710.
Fenrick R
Ballard D W
Hauber J
Böhnlein E
Cullen B R
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Article Info
Journal
Journal of virology
Abbr.
J Virol
ISSN
0022-538X
DOI
10.1128/JVI.63.8.3213-3219.1989
Published
1989-08-00
Pages
3213-9
Language
English
Region
United States
NLM ID
0113724
PMCID
PMC250890
Subset
IM
External Links
PubMed source DOI: 10.1128/JVI.63.8.3213-3219.1989 PMC Full Text View journal details
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