TRANSCRIPTOME 2002: From Functional Genomics to Systems
Gene Expression Studies of In Vitro HIV-1 Infection:HIV-1 Activates Transcription of the Sterol Biosynthesis Pathway
Angelique B. van 't Wout, Ginger K. Lehrman, Svetlana A. Mikheeva, Gemma O’Keeffe, Gary K. Geiss, Roger E. Bumgarner and James I. Mullins. University of Washington, Seattle, WA
HIV requires host cell factors to survive and replicate and needs to interfere with cellular pathways aimed at destroying viral replication. We used cDNA microarrays to assess the interplay between the various biochemical processes involved in HIV infection of T cells. The expression levels for ~4600 human cDNAs were assessed in mock versus HIV infected T cells at different times after infection. Using human T cell lines and the lab strain HIV BRU we simultaneously infected millions of cells and studied subsequent changes in gene expression levels. Some changes induced by HIV BRU were found only in certain cell types. A subset of the regulated genes was verified by RT-PCR or FACS analysis. Genes involved in transcription, translations, splicing and protein processing, including MYC, were inhibited by HIV BRU infection. Several genes involved in sterol synthesis were stimulated in infected cells. Previous work by other laboratories has shown that cholesterol is essential for HIV infectivity. Our studies suggest that HIV actively ensures sufficient levels of cholesterol in the cell membrane for release of infectious viral particles. We are currently verifying these observations using metabolic labeling to measure functional changes in the sterol synthesis pathway.
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