Phosphorylation of MdmX by CDK2/Cdc2(p34) is required for nuclear export of Mdm2.
Mdm2 and MdmX function as cellular regulators of the p53 tumor suppressor protein. Intriguingly, the activities of these proteins are interdependent; MdmX stabilizes Mdm2, enabling its activities towards p53, but it also requires Mdm2 for its nuclear localization. Here we demonstrate that via its phosphorylation by CDK2/Cdc2p34, MdmX regulates nuclear export of Mdm2. Cdc2p34 phosphorylates MdmX on Ser 96 in vitro. Mutation within this site (MdmX(S96A)) impairs, whereas phosphomimic substitution (MdmX(S96D)) increases the cytoplasmic localization of MdmX, suggesting that CDK2/Cdc2p34 phosphorylation is required for export of MdmX from the nucleus. Consequently, cells that express MdmX(S96A) retain Mdm2 in their nuclei, suggesting that export of Mdm2 to the cytoplasm is MdmX-dependent. Similarly, treatment of cells with the pharmacological inhibitor of CDK2/Cdc2p34 or with a dominant-negative Cdc2 results in nuclear localization of MdmX and Mdm2 and decreases the level of Mdm2 expression. Since Cdc2p34 is active in nonstressed conditions, our finding provides a novel insight into the signaling cascade involved in the regulation of MdmX localization and for regulation of Mdm2 localization and stability.
Elias
Bertha
B
Department of Oncological Sciences, Cancer Center, Mount Sinai School of Medicine, One Gustive L. Levy Place, Box 1130, New York, NY 10029, USA.
Laine
Aaron
A
Ronai
Ze'ev
Z
D021581:Active Transport, Cell Nucleus; D042846:CDC2-CDC28 Kinases / Q000494:pharmacology*; D002467:Cell Nucleus / Q000378:metabolism; D051357:Cyclin-Dependent Kinase 2; D003593:Cytoplasm; D006801:Humans; D009687:Nuclear Proteins / Q000378:metabolism* / Q000493:pharmacokinetics*; D010766:Phosphorylation; D011518:Proto-Oncogene Proteins / Q000378:metabolism* / Q000493:pharmacokinetics*; D051736:Proto-Oncogene Proteins c-mdm2; D015398:Signal Transduction
D009687:Nuclear Proteins; D011518:Proto-Oncogene Proteins; C497206:MDM2 protein, human; D051736:Proto-Oncogene Proteins c-mdm2; D042846:CDC2-CDC28 Kinases; C495894:CDK2 protein, human; D051357:Cyclin-Dependent Kinase 2