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SMC5/6复合物与染色质的结合依赖于其NSE3亚基与DNA的结合。

Chromatin association of the SMC5/6 complex is dependent on binding of its NSE3 subunit to DNA.

作者信息

Zabrady Katerina, Adamus Marek, Vondrova Lucie, Liao Chunyan, Skoupilova Hana, Novakova Marketa, Jurcisinova Lenka, Alt Aaron, Oliver Antony W, Lehmann Alan R, Palecek Jan J

机构信息

Mendel Centre for Plant Genomics and Proteomics, Central European Institute of Technology, Masaryk University, Kamenice 5, Brno, 62500, Czech Republic Laboratory of Functional Genomics and Proteomics, National Centre for Biomolecular Research, Faculty of Science, Masaryk University, Kotlarska 2, Brno, 61137, Czech Republic.

Mendel Centre for Plant Genomics and Proteomics, Central European Institute of Technology, Masaryk University, Kamenice 5, Brno, 62500, Czech Republic.

出版信息

Nucleic Acids Res. 2016 Feb 18;44(3):1064-79. doi: 10.1093/nar/gkv1021. Epub 2015 Oct 7.

DOI:10.1093/nar/gkv1021
PMID:26446992
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC4756808/
Abstract

SMC5/6 is a highly conserved protein complex related to cohesin and condensin, which are the key components of higher-order chromatin structures. The SMC5/6 complex is essential for proliferation in yeast and is involved in replication fork stability and processing. However, the precise mechanism of action of SMC5/6 is not known. Here we present evidence that the NSE1/NSE3/NSE4 sub-complex of SMC5/6 binds to double-stranded DNA without any preference for DNA-replication/recombination intermediates. Mutations of key basic residues within the NSE1/NSE3/NSE4 DNA-binding surface reduce binding to DNA in vitro. Their introduction into the Schizosaccharomyces pombe genome results in cell death or hypersensitivity to DNA damaging agents. Chromatin immunoprecipitation analysis of the hypomorphic nse3 DNA-binding mutant shows a reduced association of fission yeast SMC5/6 with chromatin. Based on our results, we propose a model for loading of the SMC5/6 complex onto the chromatin.

摘要

SMC5/6是一种与黏连蛋白和凝聚蛋白相关的高度保守的蛋白质复合物，而黏连蛋白和凝聚蛋白是高阶染色质结构的关键组成部分。SMC5/6复合物对于酵母增殖至关重要，并参与复制叉的稳定性和处理过程。然而，SMC5/6确切的作用机制尚不清楚。在此，我们提供证据表明，SMC5/6的NSE1/NSE3/NSE4亚复合物可结合双链DNA，且对DNA复制/重组中间体没有任何偏好。NSE1/NSE3/NSE4 DNA结合表面内关键碱性残基的突变会降低其在体外与DNA的结合。将这些突变引入粟酒裂殖酵母基因组会导致细胞死亡或对DNA损伤剂超敏。对低表达nse3 DNA结合突变体的染色质免疫沉淀分析表明，裂殖酵母SMC5/6与染色质的结合减少。基于我们的结果，我们提出了一个将SMC5/6复合物加载到染色质上的模型。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ad65/4756808/cba804478fe6/gkv1021fig7.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ad65/4756808/28225b485089/gkv1021fig1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ad65/4756808/f136bea44b16/gkv1021fig2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ad65/4756808/f020e392fb17/gkv1021fig3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ad65/4756808/5a6fd3924edf/gkv1021fig4.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ad65/4756808/55675728aeed/gkv1021fig5.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ad65/4756808/965692fb7c40/gkv1021fig6.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ad65/4756808/cba804478fe6/gkv1021fig7.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ad65/4756808/28225b485089/gkv1021fig1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ad65/4756808/f136bea44b16/gkv1021fig2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ad65/4756808/f020e392fb17/gkv1021fig3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ad65/4756808/5a6fd3924edf/gkv1021fig4.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ad65/4756808/55675728aeed/gkv1021fig5.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ad65/4756808/965692fb7c40/gkv1021fig6.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ad65/4756808/cba804478fe6/gkv1021fig7.jpg

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本文引用的文献

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Structure. 2015 Dec 1;23(12):2183-2190. doi: 10.1016/j.str.2015.10.004. Epub 2015 Nov 12.
2
The melanoma-associated antigen 1 (MAGEA1) protein stimulates the E3 ubiquitin-ligase activity of TRIM31 within a TRIM31-MAGEA1-NSE4 complex.黑色素瘤相关抗原1（MAGEA1）蛋白在TRIM31-MAGEA1-NSE4复合物中刺激TRIM31的E3泛素连接酶活性。
Cell Cycle. 2015;14(6):920-30. doi: 10.1080/15384101.2014.1000112.
3
Association of condensin with chromosomes depends on DNA binding by its HEAT-repeat subunits.
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Nucleic Acids Res. 2024 Mar 21;52(5):2112-2129. doi: 10.1093/nar/gkae103.
4
Molecular basis for Nse5-6 mediated regulation of Smc5/6 functions.Nse5-6 介导的 Smc5/6 功能调节的分子基础。
Proc Natl Acad Sci U S A. 2023 Nov 7;120(45):e2310924120. doi: 10.1073/pnas.2310924120. Epub 2023 Oct 30.
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