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ATR-dependent phosphorylation of ATRIP in response to genotoxic stress

Eisuke Itakura1, Kazuyuki Umeda, Ei Sekoguchi

  • 1Department of Geriatric Research, National Institute for Longevity Sciences, Obu, Aichi, Japan.

Biochemical and Biophysical Research Communications
|September 29, 2004
PubMed

Insights

The ATR-ATRIP complex maintains genome integrity. ATR-dependent phosphorylation of ATRIP at Ser-68 and -72 is not essential for the DNA damage response, suggesting it

Area of Science:

  • Cellular biology
  • Molecular biology
  • Genetics

Background:

  • The ATR-ATRIP complex is crucial for maintaining genome integrity by transducing DNA damage checkpoint signals.
  • The precise mechanism of ATR kinase activation upon DNA damage remains largely unknown.

Purpose of the Study:

  • To investigate the role of ATRIP phosphorylation in the ATR-dependent DNA damage response.
  • To elucidate the kinase activation mechanism of the ATR-ATRIP complex.

Main Methods:

  • In vitro kinase assays to determine ATRIP phosphorylation sites.
  • Generation of phospho-specific antibodies for phosphorylated ATRIP.
  • Analysis of ATRIP relocalization and downstream effector activation in response to genotoxic stimuli.

Main Results:

  • ATRIP is phosphorylated in an ATR-dependent manner following genotoxic stress.
  • Serine 68 and 72 residues of ATRIP are critical for its in vivo phosphorylation by ATR.
  • Phosphorylation of ATRIP is not required for its recruitment to DNA damage foci or for the activation of downstream signaling pathways.

Conclusions:

  • ATR-mediated phosphorylation of ATRIP at Ser-68 and -72 is dispensable for the initial DNA damage response.
  • The study provides new insights into the regulation of the ATR-ATRIP complex during genome surveillance.

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