Non-canonical uracil processing in DNA gives rise to double-strand breaks and deletions: relevance to class switch recombination
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- Nucl. Acids Res.-2016-Bregenhorn-2691-705
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During class switch recombination (CSR), antigen-stimulated B-cells rearrange their immunoglobulin constant heavy chain (CH) loci to generate antibodies with different effector functions. CSR is initiated by activation-induced deaminase (AID), which converts cytosines in switch (S) regions, repetitive sequences flanking the CHloci, to uracils. Although U/G mispairs arising in this way are generally efficiently repaired to C/Gs by uracil DNA glycosylase (UNG)-initiated base excision repair (BER), uracil processing in S-regions of activated B-cells occasionally gives rise to double strand breaks (DSBs), which trigger CSR. Surprisingly, genetic experiments revealed that CSR is dependent not only on AID and UNG, but also on mismatch repair (MMR). To elucidate the role of MMR in CSR, we studied the processing of uracil-containing DNA substrates in extracts of MMR-proficient and -deficient human cells, as well as in a system reconstituted from recombinant BER and MMR proteins. Here, we show that the interplay of these repair systems gives rise to DSBsin vitroand to genomic deletions and mutationsin vivo, particularly in an S-region sequence. Our findings further suggest that MMR affects pathway choice in DSB repair. Given its amenability to manipulation, our system represents a powerful tool for the molecular dissection of CSR.
Original language | English |
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Journal | Nucleic Acids Research |
Volume | 44 |
Issue number | 6 |
Pages (from-to) | 2691-2705 |
Number of pages | 15 |
ISSN | 0305-1048 |
DOIs | |
Publication status | Published - 7 Apr 2016 |
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