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DNA damage signaling recruits the Rtt107-Slx4 scaffolds via Dpb11 to mediate replication stress response

Patrice Y Ohouo1, Francisco M Bastos de Oliveira, Beatriz S Almeida

  • 1Graduate Program in Biochemistry, Molecular and Cell Biology, Weill Institute for Cell and Molecular Biology, Cornell University, 339 Weill Hall, Ithaca, New York 14853-7202, USA.

Molecular Cell
|July 31, 2010
PubMed

Insights

The DNA damage checkpoint kinase Mec1 (ATR) coordinates replication fork repair by linking Dpb11 to Slx4-Rtt107 scaffolds. Mec1-dependent phosphorylation of Slx4 is crucial for this interaction and cellular response to DNA alkylation damage.

Area of Science:

  • Molecular Biology
  • Cellular Biology
  • DNA Replication and Repair

Background:

  • The DNA damage checkpoint kinase Mec1 (ATR) is essential for maintaining replication fork integrity during cellular stress.
  • The precise mechanisms by which Mec1 regulates DNA repair factors at replication forks remain incompletely understood.

Purpose of the Study:

  • To elucidate the molecular mechanisms by which Mec1 regulates DNA repair factors at replication forks.
  • To investigate the role of Mec1 in coordinating the response to replication stress.

Main Methods:

  • Investigated protein-protein interactions using biochemical assays.
  • Utilized site-directed mutagenesis to examine the role of Mec1 phosphorylation sites.
  • Assessed cellular responses to DNA alkylation damage.

Main Results:

  • Identified a critical interaction mediated by Mec1 between Dpb11 and the Slx4-Rtt107 DNA repair scaffolds.
  • Demonstrated that Slx4 phosphorylation by Mec1 is required for its interaction with Dpb11.
  • Showed that mutations in Mec1 phosphorylation sites on Slx4 impair the cellular response to replication stress induced by DNA alkylation.

Conclusions:

  • Mec1 regulates the Slx4 and Rtt107 scaffolds, establishing a mechanistic link between DNA damage signaling and replication fork repair.
  • The Mec1-dependent assembly of the Dpb11-Slx4-Rtt107 complex is vital for coordinating multiple fork repair factors to ensure genome stability.

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