A Protein in the Yeast Saccharomyces cerevisiae Presents DNA Binding Homology to the p53 Checkpoint Protein and Tumor Suppressor

Abstract

Saccharomyces cerevisiae does not contain a p53 homolog. Utilizing this yeast as an in vivo test tube model, our aim was to investigate if a yeast protein would show p53 DNA binding homology. Electrophoretic mobility shift analyses revealed the formation of specific DNA-protein complexes consisting of S. cerevisiae nuclear protein(s) and oligonucleotides containing p53 DNA binding sites. A S. cerevisiae p53 binding site factor (Scp53BSF) bound to a p53 synthetic DNA-consensus sequence (SCS) and a p53 binding-site sequence from the MDM2 oncogene. The complexes were of comparable size. Like mammalian p53, the affinity of Scp53BSF for the SCS oligonucleotide was higher than for the MDM2 oligonucleotide. Binding of Scp53BSF to the SCS and MDM2 oligonucleotides was strongly competed by unlabeled oligonucleotides containing mammalian p53 sites, but very little by a mutated site oligonucleotide. Importantly, Scp53BSF-DNA binding activity was significantly induced in extracts from cells with DNA damage. This resulted in dose-dependent coordinated activation of transcription when using p53-binding site reporter constructs. An ancient p53-like DNA binding protein may have been found, and activation of DNA-associated factors to p53 response elements may have functions not yet determined.

Diphthamide deficiency promotes association of eEF2 with p53 to induce p21 expression and neural crest defects

“…Both S. cerevisiae and MCF7 cells are resistant to diphthamide deficiency-induced growth arrest 2,11 . Interestingly, S. cerevisiae does not have a p53 homolog 48 , and MCF7 cells contain high levels of cyclin Ds that can overcome the inhibition of cell cycle progression by p21 49 . In contrast, NC cells express substantially higher levels of p53 transcripts and are particularly susceptible to p53-mediated responses 27 .…”

High-Fidelity and Simultaneous Sensing of Endogenous Mutant and Wild p53 Proteins for Precise Cancer Diagnosis and Drug Screening

“…So far, many common methods, including enzyme-linked immunosorbent assay (ELISA), − electrophoretic mobility shift assay, , fluorescence, , immunochemistry, , and electrochemistry, , have been reported to achieve p53 protein detection. For instance, the primary antibodies in the ELISA assay were replaced by a consensus dsDNA for specific detection of Wp53 .…”

Recent Advances in p53

“…The study by Farooqi et al [ 10 ] attempted to investigate if a yeast protein would show p53 DNA binding homology. The authors present the first evidence that a protein in Saccharomyces cerevisiae has increased capability after DNA damage of recognizing and binding specifically to known p53 binding sites.…”