期刊
EMBO JOURNAL
卷 40, 期 15, 页码 -出版社
WILEY
DOI: 10.15252/embj.2021107807
关键词
chromosome segregation; cohesion; condensin; loop extrusion; Smc5; 6
资金
- Swiss National Science Foundation [310030L_170242]
- European Research Council [724482]
- German Research Foundation (DFG) [RA2941/1-1]
- Netherlands Organisation for Scientific Research (NWO) research programme TA [741.018.201]
- European Union [823839]
- Swiss National Science Foundation (SNF) [310030L_170242] Funding Source: Swiss National Science Foundation (SNF)
- European Research Council (ERC) [724482] Funding Source: European Research Council (ERC)
The Nse5/6 sub-complex inhibits the Smc5/6 ATPase by preventing productive ATP binding, which is relieved by plasmid DNA binding. This direct inhibition modulates DNA substrate selection.
Eukaryotic cells employ three SMC (structural maintenance of chromosomes) 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.
作者
我是这篇论文的作者
点击您的名字以认领此论文并将其添加到您的个人资料中。
推荐
暂无数据