TY - JOUR
T1 - An ATM- and ATR-dependent checkpoint inactivates spindle assembly by targeting CEP63
AU - Smith, Eloise
AU - Dejsuphong, Donniphat
AU - Balestrini, Alessia
AU - Hampel, Martin
AU - Lenz, Christof
AU - Takeda, Shunichi
AU - Vindigni, Alessandro
AU - Costanzo, Vincenzo
N1 - Funding Information:
We thank Tim Hunt, members of Clare Hall Laboratories and of the Genome Stability Unit for their comments. We thank H Mahbubani and J Kirk for technical support with Xenopus laevis. This work was funded by Cancer Research UK. V. Costanzo is also supported by the Lister Institute of Preventive Medicine, the European Research Council (ERC) start up grant and the EMBO Young Investigator Program (YIP). A. Vindigni thanks AIRC for its support.
PY - 2009
Y1 - 2009
N2 - Activation of the protein kinases ATM and ATR following chromosomal breakage prevents initiation of DNA replication and entry into mitosis. However, the effects of ATM and ATR activation in cells already progressing through mitosis are poorly understood. Here we report that ATM and ATR activation induced by DNA double-strand breaks (DSBs) inhibits centrosome-driven spindle assembly in Xenopus laevis mitotic egg extract and somatic cells, delaying mitotic progression. Using a cDNA expression library to screen for ATM and ATR substrates, we identified centrosomal protein CEP63 as an ATM and ATR target required for normal spindle assembly. ATM and ATR phosphorylate Xenopus CEP63 (XCEP63) on Ser 560 and promote its delocalization from the centrosome. Suppression of ATM and ATR activity or mutation of XCEP63 Ser 560 to Ala prevented spindle assembly defects. Consistently, inactivation of the CEP63 gene in avian DT40 cells impaired spindle assembly and prevented ATM- and ATR-dependent effects on mitosis. These data indicate that ATM and ATR control mitotic events in vertebrate cells by targeting CEP63 and centrosome dependent spindle assembly.
AB - Activation of the protein kinases ATM and ATR following chromosomal breakage prevents initiation of DNA replication and entry into mitosis. However, the effects of ATM and ATR activation in cells already progressing through mitosis are poorly understood. Here we report that ATM and ATR activation induced by DNA double-strand breaks (DSBs) inhibits centrosome-driven spindle assembly in Xenopus laevis mitotic egg extract and somatic cells, delaying mitotic progression. Using a cDNA expression library to screen for ATM and ATR substrates, we identified centrosomal protein CEP63 as an ATM and ATR target required for normal spindle assembly. ATM and ATR phosphorylate Xenopus CEP63 (XCEP63) on Ser 560 and promote its delocalization from the centrosome. Suppression of ATM and ATR activity or mutation of XCEP63 Ser 560 to Ala prevented spindle assembly defects. Consistently, inactivation of the CEP63 gene in avian DT40 cells impaired spindle assembly and prevented ATM- and ATR-dependent effects on mitosis. These data indicate that ATM and ATR control mitotic events in vertebrate cells by targeting CEP63 and centrosome dependent spindle assembly.
UR - http://www.scopus.com/inward/record.url?scp=61849160433&partnerID=8YFLogxK
U2 - 10.1038/ncb1835
DO - 10.1038/ncb1835
M3 - Article
C2 - 19182792
AN - SCOPUS:61849160433
SN - 1465-7392
VL - 11
SP - 278
EP - 285
JO - Nature Cell Biology
JF - Nature Cell Biology
IS - 3
ER -