Alternative end-joining catalyzes robust IgH locus deletions and translocations in the combined absence of ligase 4 and Ku70

Proc Natl Acad Sci U S A. 2010 Feb 16;107(7):3034-9. doi: 10.1073/pnas.0915067107. Epub 2010 Jan 25.

Abstract

Class switch recombination (CSR) in B lymphocytes is initiated by introduction of multiple DNA double-strand breaks (DSBs) into switch (S) regions that flank immunoglobulin heavy chain (IgH) constant region exons. CSR is completed by joining a DSB in the donor S mu to a DSB in a downstream acceptor S region (e.g., S gamma1) by end-joining. In normal cells, many CSR junctions are mediated by classical nonhomologous end-joining (C-NHEJ), which employs the Ku70/80 complex for DSB recognition and XRCC4/DNA ligase 4 for ligation. Alternative end-joining (A-EJ) mediates CSR, at reduced levels, in the absence of C-NHEJ, even in combined absence of Ku70 and ligase 4, demonstrating an A-EJ pathway totally distinct from C-NHEJ. Multiple DSBs are introduced into S mu during CSR, with some being rejoined or joined to each other to generate internal switch deletions (ISDs). In addition, S-region DSBs can be joined to other chromosomes to generate translocations, the level of which is increased by absence of a single C-NHEJ component (e.g., XRCC4). We asked whether ISD and S-region translocations occur in the complete absence of C-NHEJ (e.g., in Ku70/ligase 4 double-deficient B cells). We found, unexpectedly, that B-cell activation for CSR generates substantial ISD in both S mu and S gamma1 and that ISD in both is greatly increased by the absence of C-NHEJ. IgH chromosomal translocations to the c-myc oncogene also are augmented in the combined absence of Ku70 and ligase 4. We discuss the implications of these findings for A-EJ in normal and abnormal DSB repair.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antigens, Nuclear / genetics
  • B-Lymphocytes / immunology
  • Base Sequence
  • Blotting, Southern
  • DNA Breaks, Double-Stranded*
  • DNA Ligase ATP
  • DNA Ligases / genetics
  • DNA Primers / genetics
  • DNA Repair / immunology*
  • DNA-Binding Proteins / genetics
  • Genes, myc / genetics
  • Immunoglobulin Class Switching / genetics*
  • Immunoglobulin Class Switching / immunology
  • Immunoglobulin Heavy Chains / genetics
  • Immunoglobulin Switch Region / genetics*
  • In Situ Hybridization, Fluorescence
  • Ku Autoantigen
  • Mice
  • Mice, Knockout
  • Molecular Sequence Data
  • Translocation, Genetic / genetics*
  • Translocation, Genetic / immunology

Substances

  • Antigens, Nuclear
  • DNA Primers
  • DNA-Binding Proteins
  • Immunoglobulin Heavy Chains
  • Xrcc6 protein, mouse
  • Ku Autoantigen
  • DNA Ligases
  • DNA Ligase ATP