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pubmed-article:17142237pubmed:abstractTextActivation-induced cytidine deaminase (AID) likely initiates immunoglobulin gene-conversion (GC) by deaminating cytidines within the V-region of chicken B-cells. However, the intervening DNA lesion required to initiate GC remains elusive. GC could be initiated by a single strand break or a double strand break (DSB). To distinguish between these possibilities, we examined GC in the chicken DT40 B cell line deficient in non-homologous end joining (NHEJ). It is known that the NHEJ and homologous recombination DNA repair pathways compete for DSBs. In light of this, if a DSB is the major intermediate, deficiency in NHEJ should result in increased levels of GC. Here we show that DNA-PKcs(-/-/-) and Ku70(-/-) DT40 cells had 5- to 10-fold higher levels of GC relative to wildtype DT40 as measured by surface IgM reversion and sequencing of the V-region. These data suggest that a DSB is the major DNA lesion that initiates GC.lld:pubmed
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pubmed-article:17142237pubmed:dateRevised2009-11-19lld:pubmed
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pubmed-article:17142237pubmed:articleTitleNHEJ-deficient DT40 cells have increased levels of immunoglobulin gene conversion: evidence for a double strand break intermediate.lld:pubmed
pubmed-article:17142237pubmed:affiliationDepartment of Immunology, 5265 Medical Sciences Building, University of Toronto, Ontario, Toronto, Canada M5S 1A8.lld:pubmed
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pubmed-article:17142237pubmed:publicationTypeResearch Support, Non-U.S. Gov'tlld:pubmed
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