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HUWE1-dependent DNA-PKcs neddylation modulates its autophosphorylation in DNA damage response

Zongpei Guo1, Shaozheng Wang1, Ying Xie1

  • 1Department of Radiation Toxicology and Oncology, Beijing Key Laboratory for Radiobiology, Beijing Institute of Radiation Medicine, Beijing, 100850, P. R. China.

Cell Death & Disease
|May 28, 2020
PubMed

Insights

The study reveals that DNA-PKcs is poly-neddylated, a process regulated by HUWE1. This neddylation impacts DNA-PKcs activity and DNA repair efficiency, offering new insights into DNA damage response mechanisms.

Area of Science:

  • Molecular Biology
  • Biochemistry
  • Genetics

Background:

  • DNA-PKcs is central to DNA double-strand break repair via NHEJ.
  • DNA-PKcs activity is regulated by phosphorylation.
  • The role of NEDD8-mediated neddylation in DNA-PKcs function is unclear.

Purpose of the Study:

  • To investigate whether and how NEDD8-related neddylation affects DNA-PKcs and the NHEJ process.
  • To identify the enzymes responsible for DNA-PKcs neddylation.
  • To determine the functional consequences of DNA-PKcs neddylation on DNA repair.

Main Methods:

  • Demonstration of DNA-PKcs poly-neddylation at the kinase domain.
  • Identification of UBE2M and HUWE1 as key enzymes in DNA-PKcs neddylation.
  • Assessment of the impact of inhibiting HUWE1-dependent neddylation on DNA-PKcs autophosphorylation and NHEJ efficiency.

Main Results:

  • DNA-PKcs undergoes poly-neddylation within its kinase domain.
  • UBE2M and the E3 ligase HUWE1 are responsible for this neddylation.
  • Inhibition of HUWE1-mediated neddylation impairs DNA-PKcs autophosphorylation at Ser2056.
  • Depletion of HUWE1-dependent DNA-PKcs neddylation reduces NHEJ efficiency.

Conclusions:

  • Neddylation is a novel regulatory mechanism for DNA-PKcs.
  • HUWE1-dependent neddylation modulates DNA-PKcs activity and DNA double-strand break repair.
  • These findings provide insights into how neddylation regulates the NHEJ core complex.

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