Microinjection of wild-type adeno-associated virus type 2 (AAV-2) DNA and infectious human immunodeficiency virus type 1 (HIV-1) proviral DNA into the nuclei of human epithelioid SW480 cells leads to specific inhibition of HIV-1 replication. Mutational analysis of the AAV genome showed that this negative interference can be assigned to a functional AAV-2 gene. Moreover, the p78/p68 proteins are sufficient for the anti-HIV-1 effects. The gene also inhibits the expression of a chloramphenicol acetyltransferase (CAT) gene driven by the U3/R portion of the HIV-1 long terminal repeat (LTR) in the absence of expression. This suggests that the U3/R portion of HIV-1 contains elements responsible for the AAV-2 -mediated inhibition of HIV-1 LTR-driven CAT gene expression and, probably, also of HIV-1 replication. The results add support for the general significance of AAV-2 and specifically the gene as tools for down-regulating heterologous gene expression.


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