TRANSCRIPTIONAL REPRESSION IN NORMAL HUMAN KERATINOCYTES BY WILD-TYPE AND MUTANT P53

Wild-type p53 is a nuclear phosphoprotein that inhibits cell proliferation and represses transcriptionally most TATA box-containing promoters in transformed or tumor-derived cell lines. This study demonstrates that p53 alters transcription of the long control region (LCR) of human papillomavirus type 18 (HPV-18), Wild-type and mutant p53 143(Val to Ala) repressed the HPV-18 LCR promoter in normal human keratinocytes, the natural host cell for HPV infections. Repression by wild-type p53 was also observed in C-33A cells and in an HPV-16-immortalized cell line with an inducible wild-type p53, However, when C-33A cells were cotransfected with the HPV-18 LCR and mutant 143(Val to Ala), repression did not occur. Mutant p53 135(Cys to Ser) did not induce repression in either normal human keratinocytes or in the C-33A line; although like 143(Val to Ala), it is thought to affect the DNA binding activity of the wild-type protein. The ability of mutant p53 143(Val to Ala) to inactivate the HPV early promoter in normal cells (by approximately 60% reduction) suggests that this mutant may be able to associate with wild-type p53 and interact with TATA box-binding proteins, Therefore, these results demonstrate that the transcriptional activities of p53 mutants may be dependent upon the cell type assayed and the form of its endogenous p53, Furthermore, normal human keratinocytes represent an alternative model for determining the activities of p53 mutants.