Sae2 antagonizes Rad9 accumulation at DNA double-strand breaks to attenuate checkpoint signaling and facilitate end resection.

Abstract

The Mre11-Rad50-Xrs2^NBS1 complex plays important roles in the DNA damage response by activating the Tel1^ATM kinase and catalyzing 5'-3' resection at DNA double-strand breaks (DSBs). To initiate resection, Mre11 endonuclease nicks the 5' strands at DSB ends in a reaction stimulated by Sae2^CtIP Accordingly, Mre11-nuclease deficient (<i>mre11-nd</i>) and <i>sae2Δ</i> mutants are expected to exhibit similar phenotypes; however, we found several notable differences. First, <i>sae2Δ</i> cells exhibit greater sensitivity to genotoxins than <i>mre11-nd</i> cells. Second, <i>sae2Δ</i> is synthetic lethal with <i>sgs1Δ</i>, whereas the <i>mre11-nd sgs1Δ</i> mutant is viable. Third, Sae2 attenuates the Tel1-Rad53^CHK2 checkpoint and antagonizes Rad9^53BP1 accumulation at DSBs independent of Mre11 nuclease. We show that Sae2 competes with other Tel1 substrates, thus reducing Rad9 binding to chromatin and to Rad53. We suggest that persistent Sae2 binding at DSBs in the <i>mre11-nd</i> mutant counteracts the inhibitory effects of Rad9 and Rad53 on Exo1 and Dna2-Sgs1-mediated resection, accounting for the different phenotypes conferred by <i>mre11-nd</i> and <i>sae2Δ</i> mutations. Collectively, these data show a resection initiation independent role for Sae2 at DSBs by modulating the DNA damage checkpoint.