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Replication protein A (RPA) sumoylation positively influences the DNA damage checkpoint response in yeast

Nalini Dhingra1, Lei Wei1, Xiaolan Zhao2

  • 1From the Molecular Biology Program, Memorial Sloan Kettering Cancer Center, New York, New York 10065.

The Journal of Biological Chemistry
|December 29, 2018
PubMed

Insights

Sumoylation of replication protein A (RPA) enhances the DNA damage checkpoint. This modification, by influencing interactions with checkpoint proteins, boosts cellular survival during genotoxic stress.

Area of Science:

  • Cellular biology
  • Molecular genetics
  • Biochemistry

Background:

  • The DNA damage response (DDR) involves protein modifications like phosphorylation and sumoylation to manage genotoxic stress.
  • Sumoylation is crucial for optimal DNA damage checkpoint function, but its precise mechanisms remain unclear.
  • Replication protein A (RPA) is an early responder to DNA lesions, making it a key target for studying DDR regulation.

Purpose of the Study:

  • To investigate the role and mechanisms of RPA sumoylation in the DNA damage response in budding yeast (Saccharomyces cerevisiae).
  • To elucidate how sumoylation of RPA influences DNA damage checkpoint activation and cellular survival.

Main Methods:

  • Mapping of sumoylation sites on the large subunit of RPA (Rfa1).
  • Utilizing a sumoylation-defective Rfa1 mutant to assess the functional impact of RPA sumoylation.
  • Investigating the interaction between sumoylated Rfa1 and checkpoint proteins like Sgs1.

Main Results:

  • Sumoylation sites on Rfa1 were delineated.
  • Rfa1 sumoylation was found to operate in parallel with the 9-1-1 checkpoint complex, enhancing the DNA damage checkpoint.
  • Sumoylated Rfa1 promotes interactions with Sgs1, contributing to checkpoint kinase activation.

Conclusions:

  • SUMO modification of RPA, a DNA damage sensor, positively impacts the DNA damage checkpoint response.
  • RPA sumoylation enhances genotoxin survival by facilitating checkpoint activation and protein interactions.
Keywords:
DNA damageDNA damage responsesmall ubiquitin-like modifier (SUMO)checkpoint controlSaccharomyces cerevisiaeDNA damage checkpointDNA damage induced sumoylationRPAsumoylation, Sgs1, checkpoint kinase activation

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