Phosphorylation regulates human polη stability and damage bypass throughout the cell cycle

Nucleic Acids Res. 2017 Sep 19;45(16):9441-9454. doi: 10.1093/nar/gkx619.

Abstract

DNA translesion synthesis (TLS) is a crucial damage tolerance pathway that oversees the completion of DNA replication in the presence of DNA damage. TLS polymerases are capable of bypassing a distorted template but they are generally considered inaccurate and they need to be tightly regulated. We have previously shown that polη is phosphorylated on Serine 601 after DNA damage and we have demonstrated that this modification is important for efficient damage bypass. Here we report that polη is also phosphorylated by CDK2, in the absence of damage, in a cell cycle-dependent manner and we identify serine 687 as an important residue targeted by the kinase. We discover that phosphorylation on serine 687 regulates the stability of the polymerase during the cell cycle, allowing it to accumulate in late S and G2 when productive TLS is critical for cell survival. Furthermore, we show that alongside the phosphorylation of S601, the phosphorylation of S687 and S510, S512 and/or S514 are important for damage bypass and cell survival after UV irradiation. Taken together our results provide new insights into how cells can, at different times, modulate DNA TLS for improved cell survival.

MeSH terms

  • Cell Cycle / physiology*
  • Cell Cycle / radiation effects
  • Cell Line
  • Cell Survival
  • Cyclin-Dependent Kinase 2 / metabolism
  • DNA Damage / radiation effects
  • DNA Repair
  • DNA-Directed DNA Polymerase / chemistry
  • DNA-Directed DNA Polymerase / genetics
  • DNA-Directed DNA Polymerase / metabolism*
  • Humans
  • Phosphorylation
  • Protein Stability
  • Serine / metabolism
  • Ultraviolet Rays

Substances

  • Serine
  • CDK2 protein, human
  • Cyclin-Dependent Kinase 2
  • DNA-Directed DNA Polymerase
  • Rad30 protein