Nse5/6 inhibits the Smc5/6 ATPase to facilitate DNA substrate selection
Nse5/6 inhibits the Smc5/6 ATPase to facilitate DNA substrate selection
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Nse5/6 抑制 Smc5/6 ATPase 以促进 DNA 底物选择
DOI：
10.1101/2021.02.09.430422
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发表时间：
2021
期刊：
影响因子：
--
通讯作者：
Stephan Gruber
中科院分区：
文献类型：
--
作者：
Michael Taschner;Jérôme Basquin;Barbara Steigenberger;Ingmar Schaefer;Young-Min Soh;Claire Basquin;Esben Lorentzen;Markus Räschle;Richard A. Scheltema;Stephan Gruber
真核细胞使用三种SMC复合物来控制DNA折叠和拓扑结构。Smc 5/6复合物在DNA修复和防止有害DNA连接的积累中起作用。为了阐明Smc 5/6的具体功能如何支配这些功能，我们重建了酵母全复合物。我们发现Nse 5/6亚复合物通过阻止生产性ATP结合而强烈抑制Smc 5/6 ATP酶。这种抑制被解除质粒DNA结合，但不是由短的线性DNA，而没有Nse 5/6观察到相反的效果。基于Nse 5/6晶体结构和交联质谱数据，我们在Smc 5/6上发现了Nse 5/6的两个结合位点。一个结合位点位于Smc 5/6臂，一个位于头部，后者可能对ATP水解产生抑制作用。半胱氨酸交联证明与Nse 5/6的相互作用将ATP酶结构域锚定在非生产状态，其被ATP和DNA破坏稳定。在类似的条件下，Nse 4/3/1模块从ATP酶分离。总之，我们展示了如何通过Nse 5/6直接抑制Smc 5/6 ATP酶来调节DNA底物选择。
Eukaryotic cells employ three SMC complexes to control DNA folding and topology. The Smc5/6 complex plays roles in DNA repair and in preventing the accumulation of deleterious DNA junctions. To elucidate how specific features of Smc5/6 govern these functions, we reconstituted the yeast holo-complex. We found that the Nse5/6 sub-complex strongly inhibited the Smc5/6 ATPase by preventing productive ATP binding. This inhibition was relieved by plasmid DNA binding but not by short linear DNA, while opposing effects were observed without Nse5/6. We uncovered two binding sites for Nse5/6 on Smc5/6, based on an Nse5/6 crystal structure and cross-linking mass spectrometry data. One binding site is located at the Smc5/6 arms and one at the heads, the latter likely exerting inhibitory effects on ATP hydrolysis. Cysteine cross-linking demonstrated that the interaction with Nse5/6 anchored the ATPase domains in a non-productive state, which was destabilized by ATP and DNA. Under similar conditions, the Nse4/3/1 module detached from the ATPase. Altogether, we show how DNA substrate selection is modulated by direct inhibition of the Smc5/6 ATPase by Nse5/6.
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影响因子：
3.8
作者：
F. Onoda;M. Takeda;M. Seki;D. Maeda;J. Tajima;A. Ui;H. Yagi;T. Enomoto
通讯作者：
F. Onoda;M. Takeda;M. Seki;D. Maeda;J. Tajima;A. Ui;H. Yagi;T. Enomoto
影响因子：
2.4
作者：
Bustard DE;Ball LG;Cobb JA
通讯作者：
Cobb JA
DOI：
10.1126/science.1256917
发表时间：
2014-11-21
期刊：
Science (New York, N.Y.)
影响因子：
--
作者：
Gligoris TG;Scheinost JC;Bürmann F;Petela N;Chan KL;Uluocak P;Beckouët F;Gruber S;Nasmyth K;Löwe J
通讯作者：
Löwe J
影响因子：
16
作者：
Doyle JM;Gao J;Wang J;Yang M;Potts PR
通讯作者：
Potts PR
影响因子：
7.7
作者：
Li Y;Muir KW;Bowler MW;Metz J;Haering CH;Panne D
通讯作者：
Panne D