Title: Mdm2 and MdmX as Regulators of Gene Expression
Description: Mdm2 and MdmX are RING domain proteins that bind to and inhibit p53 trans-activation functions. Moreover, Mdm2 interacts with p53 and targets it for degradation. However, Mdm2 and MdmX function beyond a simple inhibition of p53, and increasing evidence suggests functions in regulation of target gene specificity by p53 as well as influencing gene expression through other transcription factors. In this dissertation we present two studies into the regulation of p53 target genes by MdmX and Mdm2. We found that MdmX is required for the full activation of the Mdm2 gene following cellular stress, but not of other p53 targets, such as p21. The resulting deficiency in Mdm2 induction after MdmX ablation results in impaired negative feedback loop, leading to prolonged p53 half life following DNA damage. In vitro, MdmX does not stimulate p53 interaction with Mdm2 promoter DNA. MdmX does, however, inhibit the binding of p53 to DNA to a much lesser extent than Mdm2 does. Strikingly, MdmX is required for optimal p53 binding to the Mdm2 promoter in vivo.

Thus, we have described a new mechanism by which MdmX can suppress p53, which is through transcriptional activation of p53's principal negative regulator, Mdm2. PCNA is a DNA sliding clamp that is required for DNA replication and coordinates multiple aspects of DNA biology. It is reported to be both a direct activation target of p53, as well as an indirect repression target. We have examined the roles of Mdm2 and MdmX in the regulation of the PCNA gene.