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Background  Previous studies suggest that human  RAD9  (hRad9), encoding a DNA damage checkpoint molecule, which is frequently amplified in epithelial tumor cells of breast, lung, head and neck cancer, participates in regulation of the tumor suppressor p53-dependent transactivation of pro-survival  P21   WAF1  . This study examined the exact mechanism of the hRad9 function, especially through the phosphorylation of the C-terminus, in the transcription regulation of  P21   WAF1  .    Results  The transfection of phosphorylation-defective  hRAD9  mutants of C-terminus resulted in reduction of the p53-dependent  P21   WAF1   transactivation; the knockdown of total hRad9 elicited an increased  P21   WAF1   mRNA expression. Immunoprecipitation and a ChIP assay showed that hRad9 and p53 formed a complex and both were associated with two p53-consensus DNA-binding sequences in the 5' region of  P21   WAF1   gene. The association was reduced in the experiment of phosphorylation-defective  hRAD9  mutants.    Conclusion  The present study indicates the direct involvement of hRad9 in the p53-dependent  P21   WAF1   transcriptional mechanism, presumably via the phosphorylation sites, and alterations of the hRad9 pathway might therefore contribute to the perturbation of checkpoint activation in cancer cells.

Rad9 modulates the P21 WAF1 pathway by direct association with p53