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Carcinogenesis, Vol 18, 2019-2021, Copyright © 1997 by Oxford University Press

ARTICLES

Simple identification of dominant p53 mutants by a yeast functional assay

A Inga, S Cresta, P Monti, A Aprile, G Scott, A Abbondandolo, R Iggo and G Fronza
CSTA-Mutagenesis Laboratory, National Institute for Research on Cancer (IST), Genova, Italy.

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.
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