Multifaceted regulation of the sumoylation of the Sgs1 DNA helicase.
Multifaceted regulation of the sumoylation of the Sgs1 DNA helicase.
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Sgs1 DNA解旋酶聚合化的多方面调控。
DOI：
10.1016/j.jbc.2022.102092
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发表时间：
2022-07
影响因子：
4.8
通讯作者：
Xue, Xiaoyu
中科院分区：
文献类型：
--
作者：
Li, Shibai;Mutchler, Ashley;Zhu, Xinji;So, Stephen;Epps, John;Guan, Danying;Zhao, Xiaolan;Xue, Xiaoyu
同源重组通过产生联合DNA中间体如霍利迪连接来修复DNA断裂和序列缺口。将霍利迪连接溶解成线性DNA修复产物需要酵母中Sgs 1解旋酶及其在其他生物中的同源物的活性。最近的研究表明，这些保守的解旋酶的功能受到SUMO化的调节，然而，促进其SUMO化的机制还不清楚。在这里，我们采用体外sumoylation系统和细胞测定，以确定在Sgs 1 sumoylation的DNA和支架蛋白Esc 2的作用。我们表明，DNA结合增强SGS 1类小泛素化在体外。此外，我们证明了Esc 2的中间区（MR）与DNA结合活性是必需的Sgs 1类小泛素化。出乎意料的是，我们发现Esc 2-MR的sumoylation促进作用是DNA独立的，这表明该结构域的第二个功能。与我们的生物化学数据一致，我们发现Esc 2-MR结构域，如其SUMO E2结合的C-末端结构域，其特征在于在以前的研究中，是必需的精通sumoylation的Sgs 1及其辅因子，Top3和Rmi 1，在细胞中。综上所述，这些发现提供了证据，虽然DNA结合增强Sgs 1类小泛素化，Esc 2为基础的刺激这种修饰是由两个不同的域介导的。
Homologous recombination repairs DNA breaks and sequence gaps via the production of joint DNA intermediates such as Holliday junctions. Dissolving Holliday junctions into linear DNA repair products requires the activity of the Sgs1 helicase in yeast and of its homologs in other organisms. Recent studies suggest that the functions of these conserved helicases are regulated by sumoylation; however, the mechanisms that promote their sumoylation are not well understood. Here, we employed in vitro sumoylation systems and cellular assays to determine the roles of DNA and the scaffold protein Esc2 in Sgs1 sumoylation. We show that DNA binding enhances Sgs1 sumoylation in vitro. In addition, we demonstrate the Esc2’s midregion (MR) with DNA-binding activity is required for Sgs1 sumoylation. Unexpectedly, we found that the sumoylation-promoting effect of Esc2-MR is DNA independent, suggesting a second function for this domain. In agreement with our biochemical data, we found the Esc2-MR domain, like its SUMO E2-binding C-terminal domain characterized in previous studies, is required for proficient sumoylation of Sgs1 and its cofactors, Top3 and Rmi1, in cells. Taken together, these findings provide evidence that while DNA binding enhances Sgs1 sumoylation, Esc2-based stimulation of this modification is mediated by two distinct domains.
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影响因子：
14.9
作者：
Sebesta M;Urulangodi M;Stefanovie B;Szakal B;Pacesa M;Lisby M;Branzei D;Krejci L
通讯作者：
Krejci L
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4.8
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14.9
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影响因子：
10.5
作者：
Li S;Bonner JN;Wan B;So S;Mutchler A;Gonzalez L;Xue X;Zhao X
通讯作者：
Zhao X