Transduction of the human immunodeficiency virus type 1 promoter into human chromosomal DNA by adeno-associated virus: effects on promoter activity

Nahreini, P., Mathews, M. B. (July 1997) Transduction of the human immunodeficiency virus type 1 promoter into human chromosomal DNA by adeno-associated virus: effects on promoter activity. Virology, 234 (1). pp. 42-50. ISSN 0042-6822 (Print)

URL: http://www.ncbi.nlm.nih.gov/pubmed/9234945
DOI: 10.1006/viro.1997.8623

Abstract

Transcription of the human immunodeficiency virus type 1 (HIV-1) genome takes place after integration of the provirus into human chromosomal DNA. HIV transcription is known to be modulated by viral and cellular factors but the influence of flanking chromosomal sequences on proviral gene expression has not been well defined. To investigate the activity of the integrated HIV promoter, we exploited the ability of recombinant adeno-associated virus (AAV-2) to transfer and stably integrate genes into the human genome at random or site-specifically. Chimeric AAV vectors were constructed containing an HIV-CAT reporter cassette; some vectors also contained the neomycin resistance gene to facilitate the isolation of positive clones. HeLa cells were infected with recombinant AAV, in some instances together with wild-type virus as a source of AAV rep function. We isolated 25 clones of G418-resistant cells which carried the integrated HIV-CAT cassette, generally occupying unique sites that did not correspond to the AAV-specific region of chromosome 19. The HIV promoter was transcriptionally active in most of the clones. Basal promoter activity varied substantially among the clones, and its responsivity to the HIV transactivator Tat was also variable. The integrated HIV promoter was transactivated to comparable degrees by the one-exon form and two-exon form of Tat. These findings provide evidence that the transcriptional activity of the HIV promoter can be greatly influenced by the site of proviral insertion.

Item Type: Paper
Uncontrolled Keywords: Cell Line, Transformed Chloramphenicol O-Acetyltransferase/genetics Chromosomes Dna Dependovirus Gene Expression Regulation, Viral Gene Products, tat/genetics Gene Transfer Techniques Genes, Reporter Genetic Vectors HIV-1/ genetics Hela Cells Humans Promoter Regions (Genetics) Research Support, U.S. Gov't, P.H.S. Tumor Cells, Cultured Virus Integration
Subjects: diseases & disorders > viral diseases > HIV
organism description > virus > adeno associated virus
organs, tissues, organelles, cell types and functions > cell types and functions > cell types > cell line
organs, tissues, organelles, cell types and functions > cell types and functions > cell types > cell line
organs, tissues, organelles, cell types and functions > cell types and functions > cell types > cell line

bioinformatics > genomics and proteomics > genetics & nucleic acid processing > DNA, RNA structure, function, modification > chromosome
bioinformatics > genomics and proteomics > genetics & nucleic acid processing > DNA, RNA structure, function, modification > chromosomes, structure and function > chromosome

bioinformatics > genomics and proteomics > genetics & nucleic acid processing > DNA, RNA structure, function, modification > genes, structure and function > gene transfer
CSHL Authors:
Communities: CSHL labs
Depositing User: Kathleen Darby
Date: 21 July 1997
Date Deposited: 07 May 2014 17:50
Last Modified: 07 May 2014 17:50
Related URLs:
URI: http://repository.cshl.edu/id/eprint/30002

Actions (login required)

Administrator's edit/view item Administrator's edit/view item
CSHL HomeAbout CSHLResearchEducationNews & FeaturesCampus & Public EventsCareersGiving