Cell cycle-dependent localization of CHK2 at centrosomes during mitosis

Cell Division

Estelle Schmitt²  Francis Rodier¹  Julie Lafontaine³  Isabelle Clément³  Guillaume Chouinard³ 

[1] Département de radiologie, radio-oncologie et médecine nucléaire, Université de Montréal, Montréal, Québec, Canada;Département de médecine, Université de Montréal, Montréal, Québec, Canada;Centre de recherche, Centre hospitalier de l'Université de Montréal (CRCHUM), Hôpital Notre-Dame et Institut du cancer de Montréal, Montréal, Québec, Canada

关键词: Cell cycle;    Mitosis;    Centrosome;    CHK1;    CHK2;   

Others  :  790663
DOI  :  10.1186/1747-1028-8-7

 received in 2012-12-21, accepted in 2013-05-09,  发布年份 2013

【 摘 要 】

Background

Centrosomes function primarily as microtubule-organizing centres and play a crucial role during mitosis by organizing the bipolar spindle. In addition to this function, centrosomes act as reaction centers where numerous key regulators meet to control cell cycle progression. One of these factors involved in genome stability, the checkpoint kinase CHK2, was shown to localize at centrosomes throughout the cell cycle.

Results

Here, we show that CHK2 only localizes to centrosomes during mitosis. Using wild-type and CHK2^−/− HCT116 human colon cancer cells and human osteosarcoma U2OS cells depleted for CHK2 with small hairpin RNAs we show that several CHK2 antibodies are non-specific and cross-react with an unknown centrosomal protein(s) by immunofluorescence. To characterize the localization of CHK2, we generated cells expressing inducible GFP-CHK2 and Flag-CHK2 fusion proteins. We show that CHK2 localizes to the nucleus in interphase cells but that a fraction of CHK2 associates with the centrosomes in a Polo-like kinase 1-dependent manner during mitosis, from early mitotic stages until cytokinesis.

Conclusion

Our findings demonstrate that a subpopulation of CHK2 localizes at the centrosomes in mitotic cells but not in interphase. These results are consistent with previous reports supporting a role for CHK2 in the bipolar spindle formation and the timely progression of mitosis.

【 授权许可】

   
2013 Chouinard et al.; licensee BioMed Central Ltd.

【 图 表 】

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