Ectopic restriction of DNA repair reveals that UNG2 excises AID-induced uracils predominantly or exclusively during G1 phase

J Exp Med. 2012 May 7;209(5):965-74. doi: 10.1084/jem.20112379. Epub 2012 Apr 23.

Abstract

Immunoglobulin (Ig) affinity maturation requires the enzyme AID, which converts cytosines (C) in Ig genes into uracils (U). This alone produces C:G to T:A transition mutations. Processing of U:G base pairs via U N-glycosylase 2 (UNG2) or MutSα generates further point mutations, predominantly at G:C or A:T base pairs, respectively, but it is unclear why processing is mutagenic. We aimed to test whether the cell cycle phase of U processing determines fidelity. Accordingly, we ectopically restricted UNG2 activity in vivo to predefined cell cycle phases by fusing a UNG2 inhibitor peptide to cell cycle-regulated degradation motifs. We found that excision of AID-induced U by UNG2 occurs predominantly during G1 phase, inducing faithful repair, mutagenic processing, and class switching. Surprisingly, UNG2 does not appear to process U:G base pairs at all in Ig genes outside G1 phase.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • AICDA (Activation-Induced Cytidine Deaminase)
  • Animals
  • Cytidine Deaminase / metabolism*
  • DNA Glycosylases / metabolism*
  • DNA Repair / physiology*
  • G1 Phase / physiology*
  • Genes, Immunoglobulin / genetics*
  • Genes, Immunoglobulin / physiology
  • Humans
  • Immunoglobulin Class Switching / physiology*
  • Mice
  • Mice, Inbred C57BL
  • Plasmids / genetics
  • Recombinant Fusion Proteins / metabolism
  • Time-Lapse Imaging
  • Transduction, Genetic
  • Uracil Nucleotides / metabolism

Substances

  • Recombinant Fusion Proteins
  • Uracil Nucleotides
  • CCNO protein, human
  • DNA Glycosylases
  • AICDA (Activation-Induced Cytidine Deaminase)
  • Cytidine Deaminase