Department of Biology, Rosenstiel Basic Medical Sciences Center, Brandeis University, Waltham, Massachusetts, USA.
Mol Cell Biol. 2012 Nov;32(22):4727-40. doi: 10.1128/MCB.00566-12. Epub 2012 Sep 24.
Fun30 is a Swi2/Snf2 homolog in budding yeast that has been shown to remodel chromatin both in vitro and in vivo. We report that Fun30 plays a key role in homologous recombination, by facilitating 5'-to-3' resection of double-strand break (DSB) ends, apparently by facilitating exonuclease digestion of nucleosome-bound DNA adjacent to the DSB. Fun30 is recruited to an HO endonuclease-induced DSB and acts in both the Exo1-dependent and Sgs1-dependent resection pathways. Deletion of FUN30 slows the rate of 5'-to-3' resection from 4 kb/h to about 1.2 kb/h. We also found that the resection rate is reduced by DNA damage-induced phosphorylation of histone H2A-S129 (γ-H2AX) and that Fun30 interacts preferentially with nucleosomes in which H2A-S129 is not phosphorylated. Fun30 is not required for later steps in homologous recombination. Like its homolog Rdh54/Tid1, Fun30 is required to allow the adaptation of DNA damage checkpoint-arrested cells with an unrepaired DSB to resume cell cycle progression.
Fun30 是芽殖酵母中的 Swi2/Snf2 同源物,已被证明可在体外和体内重塑染色质。我们报告说 Fun30 通过促进双链断裂 (DSB) 末端的 5'到 3'切除,显然通过促进核小体结合 DNA 的外切酶消化,在同源重组中发挥关键作用,这些 DNA 紧邻 DSB。Fun30 被招募到 HO 内切酶诱导的 DSB 中,并在 Exo1 依赖性和 Sgs1 依赖性切除途径中发挥作用。FUN30 的缺失会将 5'到 3'切除的速度从 4 kb/h 降低到约 1.2 kb/h。我们还发现,DNA 损伤诱导的组蛋白 H2A-S129(γ-H2AX)磷酸化会降低切除率,并且 Fun30 与未磷酸化的 H2A-S129 的核小体优先相互作用。Fun30 不需要同源重组的后期步骤。像它的同源物 Rdh54/Tid1 一样,Fun30 是允许具有未修复 DSB 的 DNA 损伤检查点阻滞细胞适应并恢复细胞周期进程所必需的。