Involvement of protein-DNA interaction in adeno-associated virus Rep78-mediated inhibition of HIV-1.

Important note: Information in this article was accurate in 1999. The state of the art may have changed since the publication date.

Involvement of protein-DNA interaction in adeno-associated virus Rep78-mediated inhibition of HIV-1.

J Hum Virol. 1998 Nov-Dec;1(7):441-50. Unique Identifier : AIDSLINE MED/99211169
Kokorina NA; Santin AD; Li C; Hermonat PL; Department of Obstetrics and Gynecology, University of Arkansas; for Medical Sciences, Little Rock 72205, USA.

Abstract: OBJECTIVE: It has been well documented by several laboratories that adeno-associated virus (AAV) is able to inhibit HIV-1 replication and gene expression. This effect has been mapped to the AAV-encoded Rep78 protein. However, the mechanism by which Rep78 is able to inhibit HIV-1 is unclear. As Rep78 is a DNA binding transcription factor, the objective of this study was to investigate where Rep78 might bind within the HIV-1 long terminal repeat (LTR) sequences and to judge the importance of this protein-DNA interaction. STUDY DESIGN/METHODS: Rep78's binding to HIV-LTR DNA was analyzed by electrophoretic mobility shift assay (EMSA). The importance of this protein-DNA interaction was analyzed using a Rep78 mutant defective for binding HIV-LTR DNA in an assay for monitoring gene expression (chloramphenicol acetyltransferase [CAT] assay). RESULTS: The preferred site for Rep78 binding was found to be adjacent to the HIV-LTR TATA box, within nt -54 to -34 relative to the site of transcription initiation. Furthermore, a Rep78 mutant with substitutions at amino acid residues 64 and 65 which was found defective for binding HIV-LTR DNA, was also found to be defective for inhibition of tat transactivated HIV-LTR gene expression. CONCLUSION: These data strongly suggest that Rep78's DNA binding ability is important for its mechanism of inhibition. Furthermore, the TATA box region of the HIV-LTR, to which Rep78 preferentially binds, is a likely target through which the inhibition takes place.

Keywords: JOURNAL ARTICLE Comparative Study Dependovirus/GENETICS/*PHYSIOLOGY DNA-Binding Proteins/GENETICS/*METABOLISM DNA, Viral/*METABOLISM Electrophoresis, Polyacrylamide Gel Human HIV Long Terminal Repeat/*PHYSIOLOGY HIV-1/GENETICS/*PHYSIOLOGY Mutation Support, U.S. Gov't, P.H.S. Transcription Factors/PHYSIOLOGY Transcription, Genetic/*DRUG EFFECTS TATA Box/PHYSIOLOGY Viral Interference Viral Proteins/GENETICS/*METABOLISM Virus Replication/PHYSIOLOGY
990730
A9971133

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