recovery of arrested replication forks by homologous recombination is error-prone复苏逮捕复制叉的同源重组是容易出错的.pdfVIP

Recovery of Arrested Replication Forks by Homologous Recombination Is Error-Prone 1,2. 1,2. 1,2. 1,2 ´ 1,2 Ismail Iraqui , Yasmina Chekkal , Nada Jmari , Violena Pietrobon , Karine Freon , Audrey Costes1,2, Sarah A. E. Lambert1,2* 1 Institut Curie, Centre de Recherche, Orsay, France, 2 CNRS, UMR3348, Centre Universitaire, Orsay, France Abstract Homologous recombination is a universal mechanism that allows repair of DNA and provides support for DNA replication. Homologous recombination is therefore a major pathway that suppresses non-homology-mediated genome instability. Here, we report that recovery of impeded replication forks by homologous recombination is error-prone. Using a fork-arrest- based assay in fission yeast, we demonstrate that a single collapsed fork can cause mutations and large-scale genomic changes, including deletions and translocations. Fork-arrest-induced gross chromosomal rearrangements are mediated by inappropriate ectopic recombination events at the site of collapsed forks. Inverted repeats near the site of fork collapse stimulate large-scale genomic changes up to 1,500 times over spontaneous events. We also show that the high accuracy of DNA replication during S-phase is impaired by impediments to fork progression, since fork-arrest-induced mutation is due to erroneous DNA synthesis during recovery of replication forks. The mutations caused are small insertions/duplications between short tandem repeats (micro-homology) indicative of replication slippage. Our data establish that collapsed forks, but not stalled forks, recovered by homologous recombination are prone to replication slippage. The inaccuracy of DNA synthesis doe