Abstract
Microcephalin (MCPH1/BRIT1) forms ionizing radiation-induced nuclear foci (IRIF) and is required for DNA damage-responsive S and G2-M-phase checkpoints. MCPH1 contains three BRCT domains. Here we report the cloning of chicken Mcph1 (cMcph1) and functional analysis of its individual BRCT domains. Full-length cMcph1 localized to centrosomes throughout the cell cycle and formed IRIF that colocalized with γ-H2AX. The tandem C-terminal BRCT2 and BRCT3 domains of cMcph1 were necessary for IRIF formation, while the N-terminal BRCT1 was required for centrosomal localization in irradiated cells. Centrosomal targeting of cMcph1 was independent of ATM, Brca1 or Chk1. cMcph1 formed IRIF in ATM- and Brca1-deficient cells, but not in H2AX-deficient cells. Inability to form cMcph1 IRIF impaired the cellular response to DNA damage. These results suggest that the role of microcephalin in the vertebrate DNA damage response is controlled by interaction of the C-terminal BRCT domains with γ-H2AX.
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Acknowledgements
We thank Kevin Hiom, David Gillespie and Shunichi Takeda for cell lines, Andreas Merdes and Shiaw-Yih Lin for antisera and Noel Lowndes for discussion. BJC is supported by Project Grant RP/2005/7 from the Health Research Board. The work in CGM's laboratory is funded by a Science Foundation Ireland Investigator award.
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Supplementary Information accompanies the paper on the Oncogene website (http://www.nature.com/onc).
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Jeffers, L., Coull, B., Stack, S. et al. Distinct BRCT domains in Mcph1/Brit1 mediate ionizing radiation-induced focus formation and centrosomal localization. Oncogene 27, 139–144 (2008). https://doi.org/10.1038/sj.onc.1210595
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DOI: https://doi.org/10.1038/sj.onc.1210595
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