Merck

Rad52 Restrains Resection at DNA Double-Strand Break Ends in Yeast.

Molecular cell (2019-09-23)
Zhenxin Yan, Chaoyou Xue, Sandeep Kumar, J Brooks Crickard, Yang Yu, Weibin Wang, Nhung Pham, Yuxi Li, Hengyao Niu, Patrick Sung, Eric C Greene, Grzegorz Ira
RESUMEN

Rad52 is a key factor for homologous recombination (HR) in yeast. Rad52 helps assemble Rad51-ssDNA nucleoprotein filaments that catalyze DNA strand exchange, and it mediates single-strand DNA annealing. We find that Rad52 has an even earlier function in HR in restricting DNA double-stranded break ends resection that generates 3' single-stranded DNA (ssDNA) tails. In fission yeast, Exo1 is the primary resection nuclease, with the helicase Rqh1 playing a minor role. We demonstrate that the choice of two extensive resection pathways is regulated by Rad52. In rad52 cells, the resection rate increases from ∼3-5 kb/h up to ∼10-20 kb/h in an Rqh1-dependent manner, while Exo1 becomes dispensable. Budding yeast Rad52 similarly inhibits Sgs1-dependent resection. Single-molecule analysis with purified budding yeast proteins shows that Rad52 competes with Sgs1 for DNA end binding and inhibits Sgs1 translocation along DNA. These results identify a role for Rad52 in limiting ssDNA generated by end resection.

MATERIALES
Referencia del producto
Marca
Descripción del producto

Sigma-Aldrich
Monoclonal ANTI-FLAG® M2-Peroxidase (HRP) antibody produced in mouse, clone M2, purified immunoglobulin, buffered aqueous glycerol solution
Sigma-Aldrich
(S)-(+)-Camptothecin, ≥90% (HPLC), powder
Sigma-Aldrich
Anti-HA antibody, Mouse monoclonal, clone HA-7, purified from hybridoma cell culture