Activation-induced cytidine deaminase-initiated off-target DNA breaks are detected and resolved during S phase

J Immunol. 2012 Sep 1;189(5):2374-82. doi: 10.4049/jimmunol.1200414. Epub 2012 Jul 23.

Abstract

Activation-induced cytidine deaminase (AID) initiates DNA double-strand breaks (DSBs) in the IgH gene (Igh) to stimulate isotype class switch recombination (CSR), and widespread breaks in non-Igh (off-target) loci throughout the genome. Because the DSBs that initiate class switching occur during the G₁ phase of the cell cycle, and are repaired via end joining, CSR is considered a predominantly G₁ reaction. By contrast, AID-induced non-Igh DSBs are repaired by homologous recombination. Although little is known about the connection between the cell cycle and either induction or resolution of AID-mediated non-Igh DSBs, their repair by homologous recombination implicates post-G₁ phases. Coordination of DNA breakage and repair during the cell cycle is critical to promote normal class switching and prevent genomic instability. To understand how AID-mediated events are regulated through the cell cycle, we have investigated G₁-to-S control in AID-dependent genome-wide DSBs. We find that AID-mediated off-target DSBs, like those induced in the Igh locus, are generated during G₁. These data suggest that AID-mediated DSBs can evade G₁/S checkpoint activation and persist beyond G₁, becoming resolved during S phase. Interestingly, DSB resolution during S phase can promote not only non-Igh break repair, but also Ig CSR. Our results reveal novel cell cycle dynamics in response to AID-initiated DSBs, and suggest that the regulation of the repair of these DSBs through the cell cycle may ensure proper class switching while preventing AID-induced genomic instability.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Animals
  • B-Lymphocytes / cytology
  • B-Lymphocytes / immunology
  • B-Lymphocytes / metabolism
  • Cells, Cultured
  • Cytidine Deaminase / deficiency
  • Cytidine Deaminase / genetics
  • Cytidine Deaminase / physiology*
  • DNA Breaks, Double-Stranded*
  • DNA Repair / genetics
  • DNA Repair / immunology
  • G1 Phase / genetics
  • G1 Phase / immunology
  • Immunoglobulin Class Switching / genetics*
  • Immunoglobulin Isotypes / genetics*
  • Mice
  • Mice, Inbred C57BL
  • Mice, Knockout
  • S Phase / genetics*
  • S Phase / immunology*

Substances

  • Immunoglobulin Isotypes
  • AICDA (activation-induced cytidine deaminase)
  • Cytidine Deaminase