6th International AIDS Conference


San Francisco, California, USA — June 20-23, 1990

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Identification of inhibitory sequences within the HIV-1 genome.

Int Conf AIDS 1990 Jun 20-23; 6:323 (abstract no. 1036)
Maldarelli F, Martin MA, Strebel K; Laboratory of Molecular Microbiology, NIAID, NIH, Bethesda, MD USA

OBJECTIVE: To demonstrate the existence of cis-acting, negative regulatory elements on the HIV-1 genome as a prerequisite for rev gene function.

METHODS: A novel, feedback-stimulated, HIV-1 reporter plasmid, pTS-7, was constructed using HIV-1 LTR-driven tat as a test gene. Portions of the HIV-1 genome were introduced into this test system and analyzed in transient assays for the presence of sequences which inhibit tat expression. S1 nuclease digestion assays were employed to quantitate reporter-specific RNA present in transfected cells. Putative inhibitory sequences were introduced into a second reporter plasmid, pCMV-CAT, to verify the results in an heterologous system.

RESULTS: At least two specific negative regulatory sequences were identified within the HIV-1 gag/pol gene; introduction of either element into pTS-7 resulted in an 8-10-fold reduction in expression as compared to that obtained from pTS-7. The addition of a splice donor did not abrogate the negative effect.

CONCLUSION: The HIV-1 genome contains at least two distinct, inhibitory elements within the gag/pol region which suppress gene expression.

Keywords: AEGIS, HIV-1, HIV Long Terminal Repeat, Genes, pol, Genes, rev, HIV-1 Reverse Transcriptase, Plasmids, Genome, Commonwealth of Independent States, Cats, Animal, genetics, ICA6KWDaegis,hiv-1,hivlongterminalrepeat,genes,pol,genes,rev,hiv-1reversetranscriptase,plasmids,genome,commonwealthofindependentstates,cats,animal,genetics,ica6

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