Faculty Research 1990 - 1999

Regulation of mdm2 expression by p53: alternative promoters produce transcripts with nonidentical translation potential.

Document Type

Article

Publication Date

1994

Keywords

Animal, Base-Sequence, Cell-Line-Transformed, Exons, Gene-Expression-Regulation, Genes-p53, Mice, Molecular-Sequence-Data, Promoter-Regions-(Genetics), Proto-Oncogene-Proteins, RNA-Messenger, Rabbits, SUPPORT-NON-U-S-GOVT, SUPPORT-U-S-GOVT-P-H-S, Translation-Genetic, Tumor-Cells-Cultured

First Page

1739

Last Page

1749

JAX Source

Genes Dev 1994 Aug; 8(15):1739-49.

Grant

RO1CA40099/CA/NCI

Abstract

The mdm2 proto-oncogene product binds to the p53 tumor suppressor protein and inhibits its ability to trans-activate target genes. One such target gene is mdm2 itself, which is therefore considered a component of a p53 negative feedback loop. Two tandem p53-binding motifs residing within the first intron of the murine mdm2 gene confer upon it p53-mediated activation. We now report that in murine cells p53 activates an internal mdm2 promoter (P2) located near the 3' end of intron 1, resulting in mRNA whose transcription starts within exon 2. P2 is activated by p53 within artificial constructs, as well as within the context of the chromosomal mdm2 gene. Activation follows either the introduction of overexpressed wild-type p53 into cells or the induction of endogenous wild-type p53 by ionizing radiation. The upstream, constitutive (P1) mdm2 promoter is only mildly affected by p53, if at all. The p53-derived mdm2 transcripts lack exon 1 and a few nucleotides from exon 2. As the first in-frame AUG of mdm2 is located within exon 3, the two types of mdm2 transcripts should possess similar coding potentials. Nevertheless, in vitro conditions, where each of these transcripts yields a distinct translation profile, reflect the differential usage of translation initiation codons. Initiation of translation at internal AUG codons, which occurs also in vivo, gives rise to MDM2 polypeptides incapable of binding to p53. In vitro translation profiles of the various mdm2 transcripts could be manipulated by changing the amounts of input RNA. Thus, p53 can modulate both the amount and the nature of MDM2 polypeptides through activation of the internal P2 promoter.

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