Sumoylation of the Rad1 nuclease promotes DNA repair and regulates its DNA association.
Sumoylation of the Rad1 nuclease promotes DNA repair and regulates its DNA association.
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DOI：
10.1093/nar/gku300
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发表时间：
2014-06
影响因子：
14.9
通讯作者：
Zhao X
中科院分区：
文献类型：
--
作者：
Sarangi P;Bartosova Z;Altmannova V;Holland C;Chavdarova M;Lee SE;Krejci L;Zhao X
酿酒酵母Rad 1-Rad 10复合物是一种保守的结构特异性核酸内切酶，对修复多种类型的DNA损伤很重要。在募集到损伤位点后，Rad 1-Rad 10去除受损序列，使得随后的缺口填充和连接成为可能。作用于修复的中间步骤，Rad 1-Rad 10与DNA的缔合和解离可影响修复效率。我们发现，基因毒素增强的Rad 1类小泛素化发生后，核酸酶被招募到病变部位。一个单一的赖氨酸外Rad 1的核酸酶和Rad 10结合域是sumoylated在体内和体外。该位点突变为精氨酸废除了Rad 1类小泛素化，并在高剂量DNA损伤时损害Rad 1介导的修复，但维持了单个双链断裂的修复。Rad 1类小泛素化的时间和表型偏向高病变负荷点后切口类小泛素化的作用，可能会影响Rad 1从DNA的解离。事实上，生化检查表明，类小泛素化的Rad 1降低了复合物的DNA的亲和力，而不影响其他蛋白质的性质。这些发现表明了一种模型，即Rad 1的sumoylation促进其在核酸酶切割后与DNA脱离，使其能够有效地处理大量的DNA损伤。
The Saccharomyces cerevisiae Rad1-Rad10 complex is a conserved, structure-specific endonuclease important for repairing multiple types of DNA lesions. Upon recruitment to lesion sites, Rad1-Rad10 removes damaged sequences, enabling subsequent gap filling and ligation. Acting at mid-steps of repair, the association and dissociation of Rad1-Rad10 with DNA can influence repair efficiency. We show that genotoxin-enhanced Rad1 sumoylation occurs after the nuclease is recruited to lesion sites. A single lysine outside Rad1's nuclease and Rad10-binding domains is sumoylated in vivo and in vitro. Mutation of this site to arginine abolishes Rad1 sumoylation and impairs Rad1-mediated repair at high doses of DNA damage, but sustains the repair of a single double-stranded break. The timing of Rad1 sumoylation and the phenotype bias toward high lesion loads point to a post-incision role for sumoylation, possibly affecting Rad1 dissociation from DNA. Indeed, biochemical examination shows that sumoylation of Rad1 decreases the complex's affinity for DNA without affecting other protein properties. These findings suggest a model whereby sumoylation of Rad1 promotes its disengagement from DNA after nuclease cleavage, allowing it to efficiently attend to large numbers of DNA lesions.
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影响因子：
16.8
作者：
通讯作者：
--
影响因子：
14.9
作者：
Bowles M;Lally J;Fadden AJ;Mouilleron S;Hammonds T;McDonald NQ
通讯作者：
McDonald NQ
sumoylation和DNA损伤响应。
影响因子：
5.5
作者：
Cremona CA;Sarangi P;Zhao X
通讯作者：
Zhao X
影响因子：
56.9
作者：
BARDWELL, AJ;BARDWELL, L;FRIEDBERG, EC
通讯作者：
FRIEDBERG, EC
影响因子：
7.5
作者：
Friedel, Anna M.;Pike, Brietta L.;Gasser, Susan M.
通讯作者：
Gasser, Susan M.