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Regulation of ATR substrate selection by Rad17-dependent loading of Rad9 complexes onto chromatin

Lee Zou1, David Cortez, Stephen J Elledge

  • 1Verna and Marrs McLean Department of Biochemistry and Molecular Biology, Baylor College of Medicine, Houston, Texas 77030, USA.

Genes & Development
|January 19, 2002
PubMed

Insights

Human Rad17 protein recruits the Rad9 complex to chromatin after DNA damage. This interaction is crucial for DNA repair signaling, involving ATR and Hus1, to activate the checkpoint cascade.

Area of Science:

  • Cellular Biology
  • Molecular Biology
  • Genetics

Background:

  • Cells possess signaling pathways to manage DNA damage, controlling cell cycle and repair.
  • Yeast studies implicated Rad17 and the Rad1-Rad9-Hus1 complex in sensing DNA damage.

Purpose of the Study:

  • To investigate the role of human Rad17 in DNA damage response.
  • To elucidate the interaction between Rad17, Rad9 complex, and ATR in chromatin.

Main Methods:

  • Chromatin immunoprecipitation assays.
  • Analysis of protein phosphorylation and complex formation.
  • Investigating protein localization after induced DNA damage.

Main Results:

  • Human Rad17 recruits the Rad9 protein complex to chromatin post-DNA damage.
  • Rad17 binds chromatin before damage and is phosphorylated by ATR afterward.
  • Rad17 phosphorylation is not essential for Rad9 loading; Rad17 and ATR chromatin association are independent.
  • Hus1 is required for Rad17 phosphorylation, indicating the Rad1-Rad9-Hus1 complex facilitates ATR substrate recognition.

Conclusions:

  • Multiple checkpoint protein complexes independently localize to DNA damage sites.
  • These complexes interact to initiate the DNA damage checkpoint signaling cascade.
  • Rad17 acts as a crucial mediator in recruiting the Rad9 complex and facilitating ATR-mediated signaling.

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