53BP1 cooperation with the REV7-shieldin complex underpins DNA structure-specific NHEJ

Nature. 2018 Aug;560(7716):122-127. doi: 10.1038/s41586-018-0362-1. Epub 2018 Jul 25.

Abstract

53BP1 governs a specialized, context-specific branch of the classical non-homologous end joining DNA double-strand break repair pathway. Mice lacking 53bp1 (also known as Trp53bp1) are immunodeficient owing to a complete loss of immunoglobulin class-switch recombination1,2, and reduced fidelity of long-range V(D)J recombination3. The 53BP1-dependent pathway is also responsible for pathological joining events at dysfunctional telomeres4, and its unrestricted activity in Brca1-deficient cellular and tumour models causes genomic instability and oncogenesis5-7. Cells that lack core non-homologous end joining proteins are profoundly radiosensitive8, unlike 53BP1-deficient cells9,10, which suggests that 53BP1 and its co-factors act on specific DNA substrates. Here we show that 53BP1 cooperates with its downstream effector protein REV7 to promote non-homologous end joining during class-switch recombination, but REV7 is not required for 53BP1-dependent V(D)J recombination. We identify shieldin-a four-subunit putative single-stranded DNA-binding complex comprising REV7, c20orf196 (SHLD1), FAM35A (SHLD2) and FLJ26957 (SHLD3)-as the factor that explains this specificity. Shieldin is essential for REV7-dependent DNA end-protection and non-homologous end joining during class-switch recombination, and supports toxic non-homologous end joining in Brca1-deficient cells, yet is dispensable for REV7-dependent interstrand cross-link repair. The 53BP1 pathway therefore comprises distinct double-strand break repair activities within chromatin and single-stranded DNA compartments, which explains both the immunological differences between 53bp1- and Rev7- deficient mice and the context specificity of the pathway.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cell Cycle Proteins / metabolism
  • Cell Line
  • DNA / chemistry*
  • DNA / metabolism*
  • DNA Breaks, Double-Stranded
  • DNA End-Joining Repair*
  • DNA, Single-Stranded / chemistry
  • DNA, Single-Stranded / metabolism
  • DNA-Binding Proteins / metabolism
  • Female
  • Humans
  • Immunoglobulin Class Switching / genetics
  • Mad2 Proteins / deficiency
  • Mad2 Proteins / genetics
  • Mad2 Proteins / metabolism*
  • Male
  • Mice
  • Mice, Inbred C57BL
  • Multiprotein Complexes / chemistry
  • Multiprotein Complexes / metabolism*
  • Mutation
  • Tumor Suppressor p53-Binding Protein 1 / deficiency
  • Tumor Suppressor p53-Binding Protein 1 / metabolism*
  • V(D)J Recombination / genetics

Substances

  • Cell Cycle Proteins
  • DNA, Single-Stranded
  • DNA-Binding Proteins
  • Mad2 Proteins
  • Mad2l2 protein, mouse
  • Multiprotein Complexes
  • Shld1 protein, mouse
  • Shld2 protein, mouse
  • Shld3 protein, mouse
  • Trp53bp1 protein, mouse
  • Tumor Suppressor p53-Binding Protein 1
  • DNA