Enhancement of phospholipase D activity following baculovirus and adenovirus infection in Sf9 and COS-7 cells

In order to purify the human phospholipase DI (hPLD1) for analysis of its functional properties, we applied a baculovirus-based high-expression system. As expected, Sf9 cells infected with a baculovirus encoding for the hPLD1 displayed a 7.5-fold increase in PLD activity compared to uninfected cells. Sf9 cells infected with the wild-type (WT) and other recombinant baculoviruses were used as an expression control. Surprisingly, all baculoviruses tested led to a 3-5-fold increase in basal PLD activity when compared to uninfected cells. To further characterize the nature of the increased PLD activity, the influence of ADP-ribosylation factor (ARF) and phorbol 12-myristate 13-acetate (PMA) was studied. In contrast to membranes containing the hPLD1, the PLD activity in membranes from uninfected and WT-infected Sf9 cells was not stimulated by ARF. PMA did not affect the increase in PLD activity in any case. To further study whether the virus mediated increase in PLD activity is a more general phenomenon, we infected COS-7 cells with recombinant and WT adenoviruses, Only the infection with the WT adenovirus resulted in an approx. 2-fold increase in PLD activity. Our results demonstrate for the first time that a viral infection elevates the PLD activity in insect and mammalian cells. (C) 1998 Elsevier Science B.V, All rights reserved.