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• W1968493165 abstract "This paper demonstrates that Pif1 helicase works with polymerase d to promote DNA synthesis through a migrating D-loop, a mechanism used to copy tens of kilobases during repair of chromosome breaks by break-induced replication (BIR). When DNA is repaired by homologous recombination, DNA synthesis is involved in the latter stages. Two papers published in this issue of Nature now define a role for the DNA helicase Pif1 in this reaction. They show that although the initial stages of break-induced replication (BIR) can occur normally in the absence of Pif1, synthesis from a migrating D-loop intermediate is compromised. The mechanism of replication during BIR involves a unique bubble-like replication fork that results in conservative inheritance of the new genetic material, in contrast to the S-phase replication that duplicates the genome before cell division, and is inherently mutagenic. During DNA repair by homologous recombination (HR), DNA synthesis copies information from a template DNA molecule. Multiple DNA polymerases have been implicated in repair-specific DNA synthesis1,2,3, but it has remained unclear whether a DNA helicase is involved in this reaction. A good candidate DNA helicase is Pif1, an evolutionarily conserved helicase in Saccharomyces cerevisiae important for break-induced replication (BIR)4 as well as HR-dependent telomere maintenance in the absence of telomerase5 found in 10–15% of all cancers6. Pif1 has a role in DNA synthesis across hard-to-replicate sites7,8 and in lagging-strand synthesis with polymerase δ (Polδ)9,10,11. Here we provide evidence that Pif1 stimulates DNA synthesis during BIR and crossover recombination. The initial steps of BIR occur normally in Pif1-deficient cells, but Polδ recruitment and DNA synthesis are decreased, resulting in premature resolution of DNA intermediates into half-crossovers. Purified Pif1 protein strongly stimulates Polδ-mediated DNA synthesis from a D-loop made by the Rad51 recombinase. Notably, Pif1 liberates the newly synthesized strand to prevent the accumulation of topological constraint and to facilitate extensive DNA synthesis via the establishment of a migrating D-loop structure. Our results uncover a novel function of Pif1 and provide insights into the mechanism of HR." @default.
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• W1968493165 date "2013-09-11" @default.
• W1968493165 modified "2023-10-01" @default.
• W1968493165 title "Pif1 helicase and Polδ promote recombination-coupled DNA synthesis via bubble migration" @default.
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• W1968493165 doi "https://doi.org/10.1038/nature12585" @default.
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