replication and recombination factors contributing to recombination-dependent bypass of dna lesions by template switch复制和复合因素recombination-dependent旁路开关dna损伤的模板.pdfVIP

Replication and Recombination Factors Contributing to Recombination-Dependent Bypass of DNA Lesions by Template Switch 1.¤ 1,2. 1 3 1 Fabio Vanoli , Marco Fumasoni , Barnabas Szakal , Laurent Maloisel , Dana Branzei * ` 1 Fondazione IFOM, Istituto FIRC di Oncologia Molecolare, Milan, Italy, 2 Universita degli Studi di Milano, Milan, Italy, 3 CEA, DSV, iRCM, SIGRR, LRGM, and CNRS, UMR 217, Fontenay-aux-Roses, France Abstract Damage tolerance mechanisms mediating damage-bypass and gap-filling are crucial for genome integrity. A major damage tolerance pathway involves recombination and is referred to as template switch. Template switch intermediates were visualized by 2D gel electrophoresis in the proximity of replication forks as X-shaped structures involving sister chromatid junctions. The homologous recombination factor Rad51 is required for the formation/stabilization of these intermediates, but its mode of action remains to be investigated. By using a combination of genetic and physical approaches, we show that the homologous recombination factors Rad55 and Rad57, but not Rad59, are required for the formation of template switch intermediates. The replication-proficient but recombination-defective rfa1-t11 mutant is normal in triggering a checkpoint response following DNA damage but is impaired in X-structure formation. The Exo1 nuclease also has stimulatory roles in this process. The checkpoint kinase, Rad53, is required for X-molecule formation and phosphorylates Rad55 robustly in response to DNA damage. Although Rad55 phosphorylation is thought to activate recombination