Use of CDC2 from etoposide-treated cells as substrate to assay CDC25 phosphatase activity.

C. Cans 1 V Sert J de Rycke Véronique Baldin 2 B. Ducommun 3
1 TIMC-IMAG-ThEMAS - Techniques pour l'Evaluation et la Modélisation des Actions de la Santé
TIMC-IMAG - Techniques de l'Ingénierie Médicale et de la Complexité - Informatique, Mathématiques et Applications, Grenoble - UMR 5525
2 CRBM - Centre de recherche en Biologie Cellulaire
3 LBCMCP - Laboratoire de Biologie Cellulaire et Moléculaire du Contrôle de la Prolifération
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.
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Journal articles
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https://hal-cnrs.archives-ouvertes.fr/hal-02353440
Contributor : Véronique Baldin <>
Submitted on : Thursday, November 7, 2019 - 12:13:09 PM
Last modification on : Friday, November 8, 2019 - 2:44:08 PM

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  • HAL Id : hal-02353440, version 1
  • PUBMED : 10368682

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IMAG | UNIV-TLSE3 | UGA | BS | UNIV-MONTPELLIER

Citation

C. Cans, V Sert, J de Rycke, Véronique Baldin, B. Ducommun. Use of CDC2 from etoposide-treated cells as substrate to assay CDC25 phosphatase activity.. Anticancer Research, International Institute of Anticancer Research, 1970, 19 (2A), pp.1241-4. ⟨hal-02353440⟩

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