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The AAA-ATPase VCP/p97 promotes 53BP1 recruitment by removing L3MBTL1 from DNA double-strand breaks

Klara Acs1, Martijn S Luijsterburg, Leena Ackermann

  • 1Department of Cell and Molecular Biology, Karolinska Institutet, Stockholm, Sweden.

Nature Structural & Molecular Biology
|November 29, 2011
PubMed

Insights

The tumor suppressor 53BP1 accumulates at DNA damage sites via RNF8 and RNF168 ubiquitin ligases. These enzymes recruit VCP, which displaces L3MBTL1, enabling 53BP1 binding to H4K20me2 marks.

Area of Science:

  • Molecular Biology
  • Cellular Biology
  • Genetics

Background:

  • 53BP1 is a crucial human tumor suppressor that accumulates at DNA damage sites.
  • Its recruitment relies on ubiquitin ligases RNF8 and RNF168.
  • The precise role of ubiquitylation in 53BP1 recruitment, particularly concerning its H4K20me2 binding, remained unclear.

Purpose of the Study:

  • To elucidate the mechanism by which RNF8 and RNF168 facilitate 53BP1 recruitment to DNA damage sites.
  • To investigate the role of valosin-containing protein (VCP) and its cofactors in this process.
  • To understand how histone modifications and protein displacement contribute to DNA damage response.

Main Methods:

  • Utilized RIDDLE cells (lacking RNF168) to assess VCP recruitment.
  • Investigated the interaction between VCP, L3MBTL1, and H4K20me2 marks.
  • Examined the sensitivity of nematodes lacking VCP orthologs or cofactors to ionizing radiation.

Main Results:

  • RNF8-mediated ubiquitylation recruits VCP and NPL4 to double-strand break sites.
  • RNF168-deficient RIDDLE cells exhibit impaired VCP recruitment.
  • VCP's ATPase activity releases L3MBTL1 from chromatin, facilitating 53BP1 binding to H4K20me2.
  • Nematodes lacking VCP or its cofactors show high sensitivity to ionizing radiation.

Conclusions:

  • Human RNF8 and RNF168 promote VCP-mediated displacement of L3MBTL1.
  • This displacement unmasks 53BP1 chromatin binding sites at H4K20me2 marks.
  • The findings reveal a novel pathway for 53BP1 recruitment essential for DNA damage response.

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