MEK-ERK-mediated phosphorylation of Mdm2 at Ser-166 in hepatocytes. Mdm2 is activated in response to inhibited Akt signaling.
Mdm2 inactivates the tumor suppressor p53 and Akt has been shown to be a major activator of Mdm2 in many cell types. We have investigated the regulation of Mdm2 in hepatocytes. We found that growth factor-induced Ser-166 phosphorylation of Mdm2 was inhibited by the MEK inhibitors U0126 and PD98059 in HepG2 cells and in a rat liver cell line, TRL 1215. Also, bile acids and oxidative stress induced phosphorylation of Mdm2 at Ser-166 by an apparently MEK-ERK-dependent mechanism. In contrast, Ser-166 phosphorylation of Mdm2 in lung cells was mediated by Akt. Further studies revealed that phosphatidylinositol 3-kinase inhibitors LY294002 and wortmannin induced phosphorylated ERK Tyr-204 and pMdm2 Ser-166 phosphorylations in hepatocytes in culture and in rat hepatocytes in vivo. In HepG2 cells, this effect was inhibited by U0126 and PD98059. LY294002 also reduced the level of pRaf Ser-259. Furthermore, we have shown that myr-Akt-induced overexpression of pAkt suppressed the levels of pMdm2 Ser-166 in hepatocytes. These data indicate a reversed relationship between Akt and Mdm2 in hepatocytes and suggest that Akt is a negative regulator of Raf-MEK-ERK-Mdm2 in this cell type. Ser-166 phosphorylation of Mdm2 has been shown to increase its ubiquitin ligase activity and increase p53 degradation, and our data indicated an attenuated p53 response to DNA damage in hepatocytes exhibiting high levels of pMdm2 Ser-166. Taken together, our data indicate that Mdm2 phosphorylation is regulated via MEK-ERK in hepatocytes. This Mdm2 signaling might be important for the regeneration of hepatocytes after centrilobular cell death.
Malmlöf
Maria
M
Institute of Environmental Medicine, Karolinska Institutet, S-17177 Stockholm, Sweden.
Roudier
Emilie
E
Högberg
Johan
J
Stenius
Ulla
U
D000818:Animals; D002460:Cell Line; D048049:Extracellular Signal-Regulated MAP Kinases / Q000378:metabolism; D022781:Hepatocytes / Q000378:metabolism*; D006801:Humans; D020930:MAP Kinase Kinase Kinases / Q000378:metabolism; D051058:Oncogene Protein v-akt / Q000037:antagonists & inhibitors; D010766:Phosphorylation; D051736:Proto-Oncogene Proteins c-mdm2 / Q000378:metabolism*; D051381:Rats; D011916:Rats, Inbred F344; D012694:Serine; D015398:Signal Transduction; D016159:Tumor Suppressor Protein p53 / Q000378:metabolism; D044767:Ubiquitin-Protein Ligases / Q000378:metabolism
D016159:Tumor Suppressor Protein p53; D012694:Serine; C497206:MDM2 protein, human; D051736:Proto-Oncogene Proteins c-mdm2; D044767:Ubiquitin-Protein Ligases; D051058:Oncogene Protein v-akt; D048049:Extracellular Signal-Regulated MAP Kinases; D020930:MAP Kinase Kinase Kinases