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Involvement of novel autophosphorylation sites in ATM activation

Sergei V Kozlov1, Mark E Graham, Cheng Peng

  • 1The Queensland Institute of Medical Research, Post Office Royal Brisbane Hospital, Herston, Brisbane, Queensland, Australia.

The EMBO Journal
|July 22, 2006
PubMed

Insights

ATM kinase autophosphorylation at S367 and S1893, alongside S1981, is crucial for DNA damage signaling. These phosphorylation events are vital for cellular response to DNA double-strand breaks and maintaining genome stability.

Area of Science:

  • Cellular Biology
  • Molecular Biology
  • Genetics

Background:

  • ATM kinase is essential for DNA double-strand break (DSB) signaling.
  • ATM activation requires the Mre11 complex, S1981 autophosphorylation, and other factors, but the full mechanism is unclear.

Purpose of the Study:

  • To identify and characterize additional ATM autophosphorylation sites involved in DNA damage response.
  • To investigate the physiological importance of these phosphorylation sites in vivo.

Main Methods:

  • In vitro kinase assays to assess ATM autophosphorylation.
  • Development and use of phosphospecific antibodies (anti-pS1893) for in vivo detection.
  • Analysis of ATM phosphorylation site mutants (S367A, S1893A, S1981A) in cellular models.

Main Results:

  • Identified novel ATM autophosphorylation sites at S367 and S1893, in addition to S1981.
  • Demonstrated that ATM autophosphorylates these sites in vitro upon DNA damage.
  • Observed rapid and persistent S1893 phosphorylation in vivo after ionizing radiation, dependent on ATM and Mre11.
  • Showed that phosphorylation site mutants (S367A, S1893A, S1981A) impair ATM signaling and fail to rescue DNA damage response defects.

Conclusions:

  • At least three functionally important radiation-induced autophosphorylation events occur in ATM (S367, S1893, S1981).
  • These autophosphorylation sites are critical for ATM's role in DNA damage signaling, cell cycle checkpoint control, and genome stability.

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