Inhibition of Moloney Murine Leukaemia Virus Transcription by a Phospholipase-C Inhibitor Affecting Trans -Acting Factors

Abstract

The propagation and the transcription of Moloney murine leukaemia virus (Mo-MuLV) can be inhibited by the antiviral compound tricyclodecan-9-yl-xanthate (code name: D609), which inhibits phospholipase-C (PLC) and, as a consequence, the activation of protein kinase-C (PKC) isoenzyme(s). The frans-acting factors LVa, LVb, and LVc were shown to be affected; it was not possible to retrieve them after treatment with D609 from Mo-MuLV producer cells, by virtue of the binding affinity to their consensus sequences. In contrast, the binding efficiency of the other three known transacting factors (core, NF1 and GRE), which in addition to the viral transcription, play a role in the regulation of cellular mRNA synthesis remained unimpaired. Neither LVa, LVb, nor LVc was found to be phosphorylated, which suggests that these are not targets of PKC. Only one phosphorylated DNA-binding protein was identified with an apparent molecular weight of 34kDa. This protein co-purified irrespective of the recognition sequences that we used (LVa, LVb, LVc, core, and NF1). Direct evidence is provided for the inhibition of the TPA-induced phosphorylation of the 34 kDa protein by D609. We suggest that the binding of LVa, LVb, and LVc to the DNA is mediated by the 34 kDa protein in its phosphorylated state.