Sumoylation of the Rad1 nuclease promotes DNA repair and regulates its DNA association

Details

• Alternative Title:
  Nucleic Acids Res
• Personal Author:
  Sarangi, Prabha ; Bartosova, Zdenka ; Altmannova, Veronika ; Holland, Cory ; Chavdarova, Melita ; Lee, Sang Eun ; Krejci, Lumir ; Zhao, Xiaolan
• Description:
  The Saccharomyces cerevisiae Rad1-Rad10 complex is a conserved, structure-specific endonuclease important for repairing multiple types of DNA lesions. Upon recruitment to lesion sites, Rad1-Rad10 removes damaged sequences, enabling subsequent gap filling and ligation. Acting at mid-steps of repair, the association and dissociation of Rad1-Rad10 with DNA can influence repair efficiency. We show that genotoxin-enhanced Rad1 sumoylation occurs after the nuclease is recruited to lesion sites. A single lysine outside Rad1's nuclease and Rad10-binding domains is sumoylated in vivo and in vitro. Mutation of this site to arginine abolishes Rad1 sumoylation and impairs Rad1-mediated repair at high doses of DNA damage, but sustains the repair of a single double-stranded break. The timing of Rad1 sumoylation and the phenotype bias toward high lesion loads point to a post-incision role for sumoylation, possibly affecting Rad1 dissociation from DNA. Indeed, biochemical examination shows that sumoylation of Rad1 decreases the complex's affinity for DNA without affecting other protein properties. These findings suggest a model whereby sumoylation of Rad1 promotes its disengagement from DNA after nuclease cleavage, allowing it to efficiently attend to large numbers of DNA lesions.
• Subjects:
  • Genome Integrity, Repair and Replication
  • DNA
  • DNA Damage
  • DNA Repair
  • DNA Repair Enzymes
  • Endonucleases
  • Intracellular Signaling Peptides and Proteins
  • Lysine
  • Mutation
  • Protein-Serine-Threonine Kinases
  More +
• Source:
  Nucleic Acids Res. 2014; 42(10):6393-6404.
• Pubmed ID:
  24753409
• Pubmed Central ID:
  PMC4041466
• Document Type:
  Journal Article
• Funding:
  GM071011/GM/NIGMS NIH HHS/United States ; GM080670/GM/NIGMS NIH HHS/United States ; P30 CA008748/CA/NCI NIH HHS/United States ; R01 GM071011/GM/NIGMS NIH HHS/United States ; R01 GM080670/GM/NIGMS NIH HHS/United States ; RSG-12-013-01-CCG/CC/ODCDC CDC HHS/United States
• Volume:
  42
• Issue:
  10
• Download URL:
  https://stacks.cdc.gov/view/cdc/32202/cdc_32202_DS1.pdf
• File Type:
  [PDF - 1023.89 KB]
• Collection(s):
  CDC Public Access
• Main Document Checksum:
  urn:sha256:fb20da04dbead32a03fd9ddd73c4362008556c66dd4337b8cc37db84b52cf593