The SAGA Deubiquitination Module Promotes DNA Repair and Class Switch Recombination through ATM and DNAPK-Mediated γH2AX Formation

Cell Rep. 2016 May 17;15(7):1554-1565. doi: 10.1016/j.celrep.2016.04.041. Epub 2016 May 5.

Abstract

Class switch recombination (CSR) requires activation-induced deaminase (AID) to instigate double-stranded DNA breaks at the immunoglobulin locus. DNA breaks activate the DNA damage response (DDR) by inducing phosphorylation of histone H2AX followed by non-homologous end joining (NHEJ) repair. We carried out a genome-wide screen to identify CSR factors. We found that Usp22, Eny2, and Atxn7, members of the Spt-Ada-Gcn5-acetyltransferase (SAGA) deubiquitination module, are required for deubiquitination of H2BK120ub following DNA damage, are critical for CSR, and function downstream of AID. The SAGA deubiquitinase activity was required for optimal irradiation-induced γH2AX formation, and failure to remove H2BK120ub inhibits ATM- and DNAPK-induced γH2AX formation. Consistent with this effect, these proteins were found to function upstream of various double-stranded DNA repair pathways. This report demonstrates that deubiquitination of histone H2B impacts the early stages of the DDR and is required for the DNA repair phase of CSR.

MeSH terms

  • Animals
  • Ataxia Telangiectasia Mutated Proteins / metabolism*
  • Cytidine Deaminase
  • DNA Breaks, Double-Stranded
  • DNA End-Joining Repair / radiation effects
  • DNA Repair* / radiation effects
  • DNA-Activated Protein Kinase / metabolism*
  • DNA-Binding Proteins / metabolism*
  • Endopeptidases / metabolism
  • Histones / metabolism*
  • Homologous Recombination / radiation effects
  • Immunoglobulin Class Switching / genetics*
  • Mice
  • Multiprotein Complexes / metabolism*
  • Nuclear Proteins / metabolism*
  • RNA Interference
  • Radiation, Ionizing
  • Somatic Hypermutation, Immunoglobulin / radiation effects
  • Transcription Factors / metabolism
  • Ubiquitin Thiolesterase
  • Ubiquitination* / radiation effects

Substances

  • DNA-Binding Proteins
  • Eny2 protein, mouse
  • H2AX protein, mouse
  • Histones
  • Multiprotein Complexes
  • Nuclear Proteins
  • Transcription Factors
  • Ataxia Telangiectasia Mutated Proteins
  • DNA-Activated Protein Kinase
  • Prkdc protein, mouse
  • Endopeptidases
  • USP22 protein, mouse
  • Ubiquitin Thiolesterase
  • AICDA (activation-induced cytidine deaminase)
  • Cytidine Deaminase