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Proc Natl Acad Sci U S A ; 114(4): E466-E475, 2017 01 24.
Artigo em Inglês | MEDLINE | ID: mdl-28069956

RESUMO

Cells must continuously repair inevitable DNA damage while avoiding the deleterious consequences of imprecise repair. Distinction between legitimate and illegitimate repair processes is thought to be achieved in part through differential recognition and processing of specific noncanonical DNA structures, although the mechanistic basis of discrimination remains poorly defined. Here, we show that Escherichia coli RecQ, a central DNA recombination and repair enzyme, exhibits differential processing of DNA substrates based on their geometry and structure. Through single-molecule and ensemble biophysical experiments, we elucidate how the conserved domain architecture of RecQ supports geometry-dependent shuttling and directed processing of recombination-intermediate [displacement loop (D-loop)] substrates. Our study shows that these activities together suppress illegitimate recombination in vivo, whereas unregulated duplex unwinding is detrimental for recombination precision. Based on these results, we propose a mechanism through which RecQ helicases achieve recombination precision and efficiency.


Assuntos
DNA/metabolismo , Proteínas de Escherichia coli/metabolismo , Recombinação Homóloga , RecQ Helicases/metabolismo , Reparo do DNA , Escherichia coli/enzimologia , Escherichia coli/genética , Proteínas de Escherichia coli/química , Sequências Repetidas Invertidas , RecQ Helicases/química
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