Simple identification of dominant p53 mutants by a yeast functional assay

Carcinogenesis. 1997 Oct;18(10):2019-21. doi: 10.1093/carcin/18.10.2019.

Abstract

Analysis of families with germline p53 mutations shows that the mutant p53 allele behaves as a dominant oncogene at the genetic level, although it behaves as a recessive oncogene at the cellular level, since tumours invariably show mutation or loss of both wild-type alleles. At the biochemical level it is possible that some clinically important mutant p53 proteins may be carcinogenic through a dominant mechanism. We show that p53 mutants can be readily classified according to their dominant potential using a simple yeast functional assay. Wild-type p53 is constitutively expressed from a TRP1 vector, p53 mutants are expressed from an otherwise identical LEU2 vector and net transcriptional activity is scored using an ADE2-based reporter. Twenty seven p53 mutants were tested: 19 were recessive, i.e. gave white colonies, and eight showed dominant activity, i.e. gave pink/red colonies. This simple assay should facilitate studies on p53 dominance.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • DNA, Complementary / genetics
  • Genes, Dominant / genetics*
  • Genes, Recessive / genetics
  • Genes, p53 / genetics*
  • Genetic Vectors / genetics*
  • Humans
  • Mutation / genetics*
  • Saccharomyces cerevisiae / genetics
  • Saccharomyces cerevisiae / growth & development

Substances

  • DNA, Complementary