Poly (ADP-ribose) Polymerase Is Involved in PMA-induced Activation of HIV-1 in U1 Cells by Modulating the LTR Function

Abstract

Phorbol 12-myristate 13-acetate (PMA)-induced HIV-1 production in U1 cells was markedly suppressed by inhibitors of poly (ADP-ribose) polymerase (PARP). Northern blot analysis revealed that the PARP-inhibitors suppressed the virus production at a level of transcription. In order to examine the effect of PARP on transcriptional regulation of HIV-1 genes, we transfected a reporter plasmid containing HIV-1-LTR-promoted luciferase gene to L-1210 cell clones, which expressed varying decreased level of PARP. In wild type L-1210 cells, the expression of LTR-promoted luciferase gene was stimulated approximately 4-fold in response to PMA, whereas the PMA-dependent response was almost abolished in mutant cells, which expressed only 8% of PARP of the wild type cells. The effect of decrease in PARP content on the function of HIV-1-LTR was confirmed also in human wild type cells, Jurkat and J111, which were co-transfected with the reporter plasmid and a plasmid expressing a PARP-antisense RNA: Down-regulation of PARP in the cells by the expression of the antisense RNA significantly suppressed the PMA-dependent, LTR-function of the reporter plasmid in both Jurkat and J111 cells. NF-κB, which is known to mediate the PMA-induced activation of HIV-1 in U1 cells, was found to be activated approximately 5-fold in PMA-treated U1 cells. PARP-inhibitor, unexpectedly, did not suppress but rather stimulated (approximately 2-fold) the NF-κB activation. Combining the results with the finding that the LTR-function was minimum in a PARP-defective mutant cells in spite of a very high level of the activated NF-κB in the cells, we suggest that PARP, in addition to activated NF-κB, is essential for the function of HIV-1 LTR.