ATM Phosphorylates Histone H2AX in Response to DNA Double-strand Breaks

Sandeep Burma, Benjamin P. Chen, Michael Murphy, Akihiro Kurimasa, David J. Chen

Research output: Contribution to journalArticle

1204 Citations (Scopus)

Abstract

A very early step in the response of mammalian cells to DNA double-strand breaks is the phosphorylation of histone H2AX at serine 139 at the sites of DNA damage. Although the phosphatidylinositol 3-kinases, DNA-PK (DNA-dependent protein kinase), ATM (ataxia telangiectasia mutated), and ATR (ATM and Rad3-related), have all been implicated in H2AX phosphorylation, the specific kinase involved has not yet been identified. To definitively identify the specific kinase(s) that phosphorylates H2AX in vivo, we have utilized DNA-PKcs-/- and Atm-/- cell lines and mouse embryonic fibroblasts. We find that H2AX phosphorylation and nuclear focus formation are normal in DNA-PKcs-/- cells and severely compromised in Atm-/- cells. We also find that ATM can phosphorylate H2AX in vitro and that ectopic expression of ATM in Atm-/- fibroblasts restores H2AX phosphorylation in vivo. The minimal H2AX phosphorylation in Atm-/- fibroblasts can be abolished by low concentrations of wortmannin suggesting that DNA-PK, rather than ATR, is responsible for low levels of H2AX phosphorylation in the absence of ATM. Our results clearly establish ATM as the major kinase involved in the phosphorylation of H2AX and suggest that ATM is one of the earliest kinases to be activated in the cellular response to double-strand breaks.

Original languageEnglish (US)
Pages (from-to)42462-42467
Number of pages6
JournalJournal of Biological Chemistry
Volume276
Issue number45
DOIs
StatePublished - Nov 9 2001

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Ataxia Telangiectasia
Phosphorylation
Double-Stranded DNA Breaks
Histones
DNA
Phosphotransferases
Fibroblasts
DNA-Activated Protein Kinase
Cells
Automatic teller machines
Phosphatidylinositol 3-Kinases
Serine
DNA Damage
Cell Line

ASJC Scopus subject areas

  • Biochemistry

Cite this

ATM Phosphorylates Histone H2AX in Response to DNA Double-strand Breaks. / Burma, Sandeep; Chen, Benjamin P.; Murphy, Michael; Kurimasa, Akihiro; Chen, David J.

In: Journal of Biological Chemistry, Vol. 276, No. 45, 09.11.2001, p. 42462-42467.

Research output: Contribution to journalArticle

Burma, Sandeep ; Chen, Benjamin P. ; Murphy, Michael ; Kurimasa, Akihiro ; Chen, David J. / ATM Phosphorylates Histone H2AX in Response to DNA Double-strand Breaks. In: Journal of Biological Chemistry. 2001 ; Vol. 276, No. 45. pp. 42462-42467.
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