Ig class switching.pdfVIP

nature structural molecular biology? VOLUME 18 NUMBER 1 JANUARY 2011 75 a r t i c l e s During an immune response, mature B lymphocytes undergo CSR, a deletional-recombination reaction that exchanges the Cμ constant- region gene (CH or Igh-6) of the expressed Igh for one of a set of downstream constant CH genes, such as Cγ (Ighg), Cε (Igh-7) or Cα (Igh-2). The B cell thus alters from one producing immunoglobu- lin (Ig) M to one expressing a different effector antibody molecule such as IgG, IgE or IgA1. CSR occurs between transcribed, repetitive 1–12-kb-long DNA elements termed switch (S) regions that precede each of the CH genes1. Activation-induced cytidine deaminase (AID), an essential enzyme for CSR2,3, deaminates cytidines to uridines within transcribed S regions to initiate a cascade of reactions that generates staggered DSBs4. Synapsis and end joining of DSBs between two distinct S regions completes CSR. The end-joining phase of CSR uses general DNA repair processes5. C-NHEJ, which seals DNA ends with little (1–3 nucleotides) or no homology, is a major DSB repair pathway in somatic cells6 and was thought to be essential for CSR7. However, recent studies have shown that mutations in several core C-NHEJ components including Ku70, DNA ligase IV and XRCC4 still allowed substantial CSR8–11. The mutant cells, however, had a striking alteration in the nature of the switch junctions. Although the majority of junctions in normal B cells either were blunt or had one to three base pairs of microhomology, those in the C-NHEJ mutants showed a trend toward increased micro- homology8–11. Thus, CSR proceeds through a robust A-NHEJ path- way that shows a bias toward microhomology joins. In addition to CSR, A-NHEJ has also been observed in a few other instances. First, several reporter substrates that measure joining of microhomologous DNA sequences have revealed the existence of A-NHEJ12–15. Second, although C-NHEJ is essential for end joining of DSBs g