HIV gene expression from intact proviruses positioned in bacterial artificial chromosomes at integration sites previously identified in latently infected T cells.
HIV integration predominantly occurs in introns of transcriptionally active genes. To study the impact of the integration site on HIV gene expression, a complete HIV-1 provirus (with GFP as a fusion with Nef) was inserted into bacterial artificial chromosomes (BACs) at three sites previously identified in latent T cells of patients: topoisomerase II (Top2A), DNA methyltransferase 1 (DNMT1), or basic leucine transcription factor 2 (BACH2). Transfection of BAC-HIV into 293T cells resulted in a fourfold difference in production of infectious HIV-1. Cell lines were established that contained BAC-Top2A, BAC-DNMT1, or BAC-BACH2, but only BAC-DNMT1 spontaneously produced virus, albeit at a low level. Stimulation with TNF-α resulted in virus production from four of five BAC-Top2A and all BAC-DNMT1 cell lines, but not from the BAC-BACH2 lines. The results of these studies highlight differences between integration sites identified in latent T cells to support virus production and reactivation from latency.
Department of Cell Biology, University of Alabama at Birmingham, 802 Kaul Building, 720 20th Street South, Birmingham, AL 35294, USA.
Peter G
PG
Eipers
Jesus F
JF
Salazar-Gonzalez
Casey D
CD
Morrow
D002460:Cell Line; D022202:Chromosomes, Artificial, Bacterial; D015967:Gene Expression Regulation, Viral / Q000502:physiology*; D049452:Green Fluorescent Proteins; D015497:HIV-1 / Q000235:genetics* / Q000378:metabolism*; D006801:Humans; D011533:Proviruses; D013601:T-Lymphocytes / Q000821:virology*; D016662:Virus Integration
D049452:Green Fluorescent Proteins