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Histone H2AX is a critical factor for cellular protection against DNA alkylating agents

Oncogene volume 27, pages 5662–5671 (2008)Cite this article

Abstract

Histone H2A variant H2AX is a dose-dependent suppressor of oncogenic chromosome translocations. H2AX participates in DNA double-strand break repair, but its role in other DNA repair pathways is not known. In this study, role of H2AX in cellular response to alkylation DNA damage was investigated. Cellular sensitivity to two monofunctional alkylating agents (methyl methane sulfonate and N-methyl-N′-nitro-N-nitrosoguanidine (MNNG)) was dependent on H2AX dosage, and H2AX null cells were more sensitive than heterozygous cells. In contrast to wild-type cells, H2AX-deficient cells displayed extensive apoptotic death due to a lack of cell-cycle arrest at G2/M phase. Lack of G2/M checkpoint in H2AX null cells correlated well with increased mitotic irregularities involving anaphase bridges and gross chromosomal instability. Observation of elevated poly(ADP) ribose polymerase 1 (PARP-1) cleavage suggests that MNNG-induced apoptosis occurs by PARP-1-dependent manner in H2AX-deficient cells. Consistent with this, increased activities of PARP and poly(ADP) ribose (PAR) polymer synthesis were detected in both H2AX heterozygous and null cells. Further, we demonstrate that the increased PAR synthesis and apoptotic death induced by MNNG in H2AX-deficient cells are due to impaired activation of mitogen-activated protein kinase pathway. Collectively, our novel study demonstrates that H2AX, similar to PARP-1, confers cellular protection against alkylation-induced DNA damage. Therefore, targeting either PARP-1 or histone H2AX may provide an effective way of maximizing the chemotherapeutic value of alkylating agents for cancer treatment.

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Acknowledgements

We acknowledge the generous gift of mouse ES cells proficient and deficient in H2AX from Dr CH Bassing and Dr FW Alt at the Department of Genetics, The CBR Institute for Biomedical Research, The Children's Hospital, Harvard Medical School, Boston, MA, USA. This study was partially supported by a research grant from US Department of Energy, Office of Sciences (BER) awarded to ASB (DE-FG02-05ER64055) and CRG (DE-FG 02-05ER 64054). ASB and CRG acknowledge the financial support received from NIH/NCI (5P01CA49062-16).

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Authors and Affiliations

  1. Department of Radiation Oncology, College of Physicians and Surgeons, Center for Radiological Research, Columbia University Medical Center, New York, NY, USA

    J A Meador, M Zhao, Y Su, G Narayan, C R Geard & A S Balajee

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  1. J A Meador
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  2. M Zhao
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  4. G Narayan
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Correspondence to A S Balajee.

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Supplementary Information accompanies the paper on the Oncogene website (http://www.nature.com/onc)

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Meador, J., Zhao, M., Su, Y. et al. Histone H2AX is a critical factor for cellular protection against DNA alkylating agents. Oncogene 27, 5662–5671 (2008). https://doi.org/10.1038/onc.2008.187

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