Poliovirus 3C protease-mediated degradation of transcriptional activator p53 requires a cellular activity

Abstract

Infection of HeLa cells with poliovirus leads to rapid shut-off of host cell transcription by RNA polymerase II. Previous results have suggested that both the basal transcription factor TBP (TATA-binding protein) and transcription activator proteins such as CREB (cyclic AMP-responsive element-binding protein) and Oct-1 (the octamer-binding factor) are cleaved by the viral-encoded protease, 3CPro. Here we demonstrate that the transcriptional activator (and tumor suppressor) p53 is degraded by the viral protease 3C both in vivo and in vitro. Unlike other transcription factors that are directly cleaved by 3CPro, degradation of p53 requires a HeLa cell activity in addition to 3CPro. The degradation of p53 by 3CPro does not appear to involve the ubiquitin pathway of protein degradation. Vaccinia virus infection of HeLa cells leads to inactivation of the cellular activity required for 3CPro-mediated degradation of p53. The vaccinia-encoded protein (CrmA) is known to inhibit caspase I (ICE protease) that converts inactive IL-1β to an active secreted form. Incubation of HeLa cells with caspase I inhibitor Z-VAD-fmk does not interfere with 3CPro-mediated degradation of p53. The cellular activity present in extracts of HeLa cells can be fractionated through phosphocellulose. A partially purified fraction that elutes at 0.6 M KCI from phosphocellulose contains the activity that degrades p53 in a 3CPro-dependent manner. These results suggest that both poliovirus-encoded protease 3CPro and a cellular activity are required for the degradation of p53 observed in cells infected with poliovirus. © 2001 Elsevier Science.