Phosphorylation of CDC25B by Aurora-A at the centrosome contributes to the G2-M transition.
Aurora-A protein kinase, which is the product of an oncogene, is required for the assembly of a functional mitotic apparatus and the regulation of cell ploidy. Overexpression of Aurora-A in tumour cells has been correlated with cancer susceptibility and poor prognosis. Aurora-A activity is required for the recruitment of CDK1-cyclin B1 to the centrosome prior to its activation and the commitment of the cell to mitosis. In this report, we demonstrate that the CDC25B phosphatase, an activator of cyclin dependent kinases at mitosis, is phosphorylated both in vitro and in vivo by Aurora-A on serine 353 and that this phosphorylated form of CDC25B is located at the centrosome during mitosis. Knockdown experiments by RNAi confirm that the centrosome phosphorylation of CDC25B on S353 depends on Aurora-A kinase. Microinjection of antibodies against phosphorylated S353 results in a mitotic delay whilst overexpression of a S353 phosphomimetic mutant enhances the mitotic inducing effect of CDC25B. Our results demonstrate that Aurora-A phosphorylates CDC25B in vivo at the centrosome during mitosis. This phosphorylation might locally participate in the control of the onset of mitosis. These findings re-emphasise the role of the centrosome as a functional integrator of the pathways contributing to the triggering of mitosis.
Dutertre
Stéphanie
S
Groupe Cycle Cellulaire - CNRS UMR6061 - IFR97, Génomique Fonctionnelle et Santé, Université de Rennes I, 35043 Rennes, France.
Cazales
Martine
M
Quaranta
Muriel
M
Froment
Carine
C
Trabut
Valerie
V
Dozier
Christine
C
Mirey
Gladys
G
Bouché
Jean-Pierre
JP
Theis-Febvre
Nathalie
N
Schmitt
Estelle
E
Monsarrat
Bernard
B
Prigent
Claude
C
Ducommun
Bernard
B
D000906:Antibodies / Q000378:metabolism; D000911:Antibodies, Monoclonal / Q000378:metabolism; D064127:Aurora Kinases; D018797:Cell Cycle Proteins / Q000737:chemistry / Q000378:metabolism*; D002455:Cell Division / Q000502:physiology*; D018385:Centrosome / Q000378:metabolism*; D016195:G2 Phase / Q000502:physiology*; D006367:HeLa Cells; D006801:Humans; D008845:Microinjections; D010766:Phosphorylation; D011494:Protein Kinases / Q000378:metabolism*; D017346:Protein Serine-Threonine Kinases; D034622:RNA Interference; D012694:Serine / Q000378:metabolism; D013997:Time Factors; D029867:Xenopus Proteins; D020687:cdc25 Phosphatases / Q000737:chemistry / Q000378:metabolism*
D000906:Antibodies; D000911:Antibodies, Monoclonal; D018797:Cell Cycle Proteins; D029867:Xenopus Proteins; D012694:Serine; D011494:Protein Kinases; C112134:AURKA protein, Xenopus; D064127:Aurora Kinases; D017346:Protein Serine-Threonine Kinases; C517008:CDC25B protein, human; D020687:cdc25 Phosphatases