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Journal articles
Use of CDC2 from etoposide-treated cells as substrate to assay CDC25 phosphatase activity
Abstract : Cyclin-dependent kinases (CDKs) regulate the key transition of the cell cycle in all organisms. In response to Etoposide (VP-16) induced DNA damage, cells undergo a G2-phase arrest resulting in the accumulation of inactive CDK1 (CDC2) kinase complexes. Here we report that upon Etoposide treatment CDC2 is phosphorylated on tyrosine 15 and is dephosphorylated and activated in vitro by recombinant CDC25 phosphatase. We also show that inactive CDC2 kinase from Etoposide-treated cells can be used as a substrate in a sensitive two-step assay of CDC25 phosphatase. This assay, which is very simple to set-up, is based on the monitoring of CDC2 kinase activity after CDC25-dependent dephosphorylation. It provides the possibility to use a highly physiological substrate in antimitotic drugs screening.
https://hal-cnrs.archives-ouvertes.fr/hal-02353440
Contributor : Véronique Baldin Connect in order to contact the contributor
Submitted on : Thursday, November 7, 2019 - 12:13:09 PM
Last modification on : Wednesday, October 20, 2021 - 3:44:05 AM
Contributor : Véronique Baldin Connect in order to contact the contributor
Submitted on : Thursday, November 7, 2019 - 12:13:09 PM
Last modification on : Wednesday, October 20, 2021 - 3:44:05 AM