XB-ART-8891Methods Mol Med. June 1, 2001; 12 (6): 1791-9.
The polo-like kinase Plx1 is required for activation of the phosphatase Cdc25C and cyclin B-Cdc2 in Xenopus oocytes.
In the Xenopus oocyte system mitogen treatment triggers the G(2)/M transition by transiently inhibiting the cAMP-dependent protein kinase (PKA); subsequently, other signal transduction pathways are activated, including the mitogen-activated protein kinase (MAPK) and polo-like kinase pathways. To study the interactions between these pathways, we have utilized a cell-free oocyte extract that carries out the signaling events of oocyte maturation after addition of the heat-stable inhibitor of PKA, PKI. PKI stimulated the synthesis of Mos and activation of both the MAPK pathway and the Plx1/Cdc25C/cyclin B-Cdc2 pathway. Activation of the MAPK pathway alone by glutathione S-transferase (GST)-Mos did not lead to activation of Plx1 or cyclin B-Cdc2. Inhibition of the MAPK pathway in the extract by the MEK1 inhibitor U0126 delayed, but did not prevent, activation of the Plx1 pathway, and inhibition of Mos synthesis by cycloheximide had a similar effect, suggesting that MAPK activation is the only relevant function of Mos. Immunodepletion of Plx1 completely inhibited activation of Cdc25C and cyclin B-Cdc2 by PKI, indicating that Plx1 is necessary for Cdc25C activation. In extracts containing fully activated Plx1 and Cdc25C, inhibition of cyclin B-Cdc2 by p21(Cip1) had no significant effect on either the phosphorylation of Cdc25C or the activity of Plx1. These results demonstrate that maintenance of Plx1 and Cdc25C activity during mitosis does not require cyclin B-Cdc2 activity.
PubMed ID: 11408585
PMC ID: PMC37341
Article link: Methods Mol Med.
Grant support: CA46934 NCI NIH HHS , DK28353-19 NIDDK NIH HHS , GM26743-21 NIGMS NIH HHS , R01 GM026743 NIGMS NIH HHS , R01 DK028353 NIDDK NIH HHS , CA46934 NCI NIH HHS , P30 CA046934 NCI NIH HHS , GM26743-21 NIGMS NIH HHS , DK28353-19 NIDDK NIH HHS
Genes referenced: ccnb1.2 cdc25c cdk1 cdkn1a map2k1 mapk1 mos plk1 pold1