Development of screening systems for drugs against human papillomavirus-associated cervical cancer: based on E6-E6AP binding

Human papillomavirus (HPV) E6 protein forms a ternary complex with the cell -cycle regulator p53 and the E6-associated protein (E6AP) known as an E3 ub iquitin protein ligase, leading to the degradation of p53 via the ubiquitin ation pathway. As an attempt to employ interaction between HPV viral oncoge ne E6 and a cellular protein E6AP for in vitro screening system of drugs ag ainst HPV infection, we primarily investigated the E6AP-E6 binding through Full down assay and enzyme-linked immunosorbent assay (ELISA). E6AP immobil ized on the resin produced specifically complexes with bacterially expresse d E6 in a dose-dependent manner? as determined by immunoblot analysis. This result was collinear with that shown in ELISA, which is a useful system fo r mass-screening potential drugs with rapidity and cheapness. Screening sys tem based on the interaction between E6AP and E6 may be a promising system in the development of drugs against cervical cancer caused by HPV infection . (C) 2000 Elsevier Science B.V. All rights reserved.