Wiltzius Jed J W, Hohl Marcel, Fleming James C, Petrini John H J
Molecular Biology Program, Memorial Sloan Kettering Cancer Center and Cornell University Graduate School of Medical Sciences, RRL 901B, 1275 York Avenue, New York, New York 10021, USA.
Nat Struct Mol Biol. 2005 May;12(5):403-7. doi: 10.1038/nsmb928. Epub 2005 Apr 24.
The Mre11 complex (in Saccharomyces cerevisiae: Mre11, Rad50 and Xrs2) influences multiple facets of chromosome break metabolism. A conserved feature of the Mre11 complex is a zinc-coordinating motif in Rad50 called the Rad50 hook. We established a diploid yeast strain, rad50(hook), in which Rad50 is encoded in halves, one from each of the two RAD50 alleles, with the residues constituting the hook deleted. In all respects, rad50(hook) phenocopies complete Rad50 deficiency. Replacing the hook domain with a ligand-inducible FKBP dimerization cassette partially mitigated all phenotypes in a ligand-dependent manner. The data indicate that the Rad50 hook is critical for Mre11 complex-dependent DNA repair, telomere maintenance and meiotic double-strand break formation. Sister chromatid cohesion was unaffected by Rad50 deficiency, suggesting that molecular bridging required for recombinational DNA repair is qualitatively distinct from cohesin-mediated sister chromatid cohesion.
Mre11复合物(在酿酒酵母中:Mre11、Rad50和Xrs2)影响染色体断裂代谢的多个方面。Mre11复合物的一个保守特征是Rad50中一个与锌配位的基序,称为Rad50钩。我们构建了一个二倍体酵母菌株rad50(hook),其中Rad50由两个RAD50等位基因各自编码一半,构成钩的残基被删除。在所有方面,rad50(hook)模拟了完全Rad50缺陷的表型。用配体诱导的FKBP二聚化盒替换钩结构域以配体依赖的方式部分缓解了所有表型。数据表明,Rad50钩对于Mre11复合物依赖的DNA修复、端粒维持和减数分裂双链断裂形成至关重要。姐妹染色单体黏连不受Rad50缺陷的影响,这表明重组DNA修复所需的分子桥接在性质上不同于黏连蛋白介导的姐妹染色单体黏连。