Abstract:
Replication of SIV, as well as HIV-1 and -2, requires the functional expression of the virally encoded rev protein. The rev protein acts posttranscriptionally to control the cytoplasmic expression of certain viral RNAs that are otherwise retained in the nucleus. Results of transient transfection assays have shown that trans-dominant HIV-1 rev mutants can inhibit the production of HIV-1 when cotransfected with a proviral DNA molecular clone (Malim et al., Cell 58, 1989). To define the potential use of trans-dominant rev mutants as reagents for gene therapy approaches to treat AIDS patients, we have initiated studies with SIVmac239 to evaluate their potential therapeutic value in an animal model. Because HIV-1 rev can functionally substitute for SIV rev, we used trans-dominant HIV-1 rev mutants for these studies. In our transient transfection assays, HIV-1 rev mutant, M10, which contains a two amino acid substitution within the rev activation domain, and HIV-1 rev mutants A4 and A5, which contain 4 and 5 leucine to alanine substitutions within the rev activation domain, inhibit SIVmac239 production from a full-length infectious proviral DNA molecular clone (pBRmac239) greater than 95%. As a control, HIV-1 provirus (pNL4-3) replication was also assayed and found to be inhibited by the M10, A4 and A5 rev mutants greater than 95%. To determine whether such trans-dominant rev proteins could provide long-term protection against SIV infection, we constructed retroviral vectors encoding the trans-dominant mutant rev proteins, M10 and A5, for use in transducing CEMx174 cells. Studies involving these stably transduced CEMx174 cell lines suggest that trans-dominant HIV-1 rev mutants should be further evaluated in an animal model to determine their efficacy.
Keywords: Acquired Immunodeficiency Syndrome/THERAPY Animal Cell Line Disease Models, Animal Gene Products, rev/GENETICS/*METABOLISM Gene Therapy *Genes, rev Genes, Dominant Human HIV-1/*GENETICS/PHYSIOLOGY HIV-2/PHYSIOLOGY Mutagenesis, Site-Directed RNA Processing, Post-Transcriptional RNA, Viral/BIOSYNTHESIS SIV/GENETICS/*PHYSIOLOGY Transfection Virus Replication/*GENETICS ABSTRACT 940730
M9470917
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