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Published 10 October 2005. doi:10.1083/jcb.200503066
The Rockefeller University Press, 0021-9525 $8.00
JCB, Volume 171, Number 1, 35-45
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Article

 Cdc25B cooperates with Cdc25A to induce mitosis but has a unique role in activating cyclin B1–Cdk1 at the centrosome

Arne Lindqvist, Helena Källström, Andreas Lundgren, Emad Barsoum, and Christina Karlsson Rosenthal

Department of Cell and Molecular Biology, Karolinska Institutet, S-171 77 Stockholm, Sweden

Correspondence to Christina Karlsson Rosenthal: christina.karlsson{at}cmb.ki.se

Cdc25 phosphatases are essential for the activation of mitotic cyclin–Cdks, but the precise roles of the three mammalian isoforms (A, B, and C) are unclear. Using RNA interference to reduce the expression of each Cdc25 isoform in HeLa and HEK293 cells, we observed that Cdc25A and -B are both needed for mitotic entry, whereas Cdc25C alone cannot induce mitosis. We found that the G2 delay caused by small interfering RNA to Cdc25A or -B was accompanied by reduced activities of both cyclin B1–Cdk1 and cyclin A–Cdk2 complexes and a delayed accumulation of cyclin B1 protein. Further, three-dimensional time-lapse microscopy and quantification of Cdk1 phosphorylation versus cyclin B1 levels in individual cells revealed that Cdc25A and -B exert specific functions in the initiation of mitosis: Cdc25A may play a role in chromatin condensation, whereas Cdc25B specifically activates cyclin B1–Cdk1 on centrosomes.

Abbreviations used in this paper: 3D, three-dimensional; dsRED, Discosoma red fluorescent protein; NF-Y, nuclear transcription factor Y; pCFP, plasmid CFP; RNAi, RNA interference; siRNA, small interfering RNA.


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