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Nucleic Acids Research, 2001, Vol. 29, No. 5 1222-1227
© 2001 Oxford University Press

Translational control of human p53 expression in yeast mediated by 5'-UTR–ORF structural interaction

Raja Mokdad-Gargouri*, Khmaies Belhadj and Ali Gargouri

Laboratoire ‘Génétique Moléculaire des Eucaryotes’, Centre de Biotechnologie de Sfax, BP‘K’3038, Sfax-Tunisia

We have expressed human p53 cDNA in the yeast Saccharomyces cerevisiae and shown that the level of production and the length of the p53 protein depends on the presence of untranslated mRNA regions (UTRs). The expression of the ORF alone leads to a p53 protein of correct size (53 kDa) that accumulates to high levels, concomitantly with the presence of a small amount of a p40 protein (40 kDa). However, when either the entire 5'-UTR and a part of the 3'- or 5'-UTR alone is used, this leads to the production of small amounts of the 40 kDa truncated form only. The p40 protein corresponds to a truncated form of p53 at the C-terminal extremity since it reacts only with a monoclonal antibody recognising the N-terminal epitope. This effect on the amount and length of p53 protein had no correlation at the mRNA level, suggesting that translational control probably occurs through the 5'-UTR. We propose a model of structural interaction between this UTR and a part of the ORF mRNA for the regulation of p53 expression in this heterologous context.

* To whom correspondence should be addressed. Tel: +216 4 274 110; Fax: +216 4 275 970; Email: raja.gargouri{at}cbs.rnrt.tn Present address: Khmaies Belhadj, Department of Molecular Biology and Genetics, Bilkent University, 06533 Bilkent Ankara, Turkey


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