NuRD subunit MTA1 interacts with the DNA non-homologous end joining Ku complex in cancer cells.
NuRD subunit MTA1 interacts with the DNA non-homologous end joining Ku complex in cancer cells.
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NuRD 亚基 MTA1 与癌细胞中 DNA 非同源末端连接 Ku 复合物相互作用
DOI：
10.1039/c8ra06907g
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发表时间：
2018-10-10
期刊：
影响因子：
3.9
通讯作者：
Qian, Haili
中科院分区：
文献类型：
--
作者：
Liu, Jian;Liu, Qun;Wang, Haijuan;Li, Chunxiao;Wen, Tao;An, Guangyu;Qian, Haili
转移相关抗原1（MTA 1）是一种染色质修饰剂，介导DNA修饰和基因表达。Ku 70/Ku 80复合物在DNA损伤反应中起重要作用。为了探索MTA 1相互作用组，我们在结肠癌细胞系中用两种特异性MTA 1抗体捕获了Ku 70/Ku 80复合物。我们首先通过细胞裂解物中的免疫共沉淀（co-IP）分析验证了MTA 1与Ku复合物之间的体外相互作用，表明这种相互作用主要发生在细胞核中，但也以较低水平存在于细胞质中。我们使用邻近连接测定（PLA）进一步可视化并确认了它们的体内相互作用，其与体外分析一致，也证明了绝大多数相互作用图在细胞核中，少量在细胞质中。我们以前证明，MTA 1动态和周期性分布在细胞周期。在这里，通过荧光共定位，我们发现，MTA 1和Ku蛋白共定位在细胞核中的间期和同步移动从前期到后期。有趣的是，在末期，当MTA 1被报道重新进入细胞核时，它们被分离并不同步地移动。此外，使用原位聚乳酸，我们可视化的相互作用发生在间期和有丝分裂。在间期，它们主要在细胞核内相互作用，但在有丝分裂期间，它们在染色体周围相互作用。我们还发现，MTA 1与Ku在癌组织和正常组织中均具有良好的相关性，并且它们在UV诱导的DNA损伤反应中具有协同作用。总的来说，我们的数据揭示了MTA 1和Ku复合物在细胞核和细胞质以及整个细胞周期中的特异性相互作用。因此，我们提出了一个潜在的功能之间的串扰NuRD和Ku复合物，在细胞中的两个最基本的功能单元，通过物理相互作用。
Metastasis-associated antigen 1 (MTA1) is a chromatin modifier mediating DNA modification and gene expression. Ku70/Ku80 complex has been reported to be essential in DNA damage response. In an effort to explore the MTA1 interactome, we captured the Ku70/Ku80 complex with two specific MTA1 antibodies in a colon cancer cell line. We first validated the in vitro interaction between MTA1 and the Ku complex by co-immunoprecipitation (co-IP) analyses in cell lysate, showing that the interaction occurred mainly at the nucleus, but also existed in the cytoplasm at a lower level. We further visualized and confirmed their in vivo interaction using proximity ligation assay (PLA), which, in line with the in vitro analysis, also demonstrated a vast majority of interaction plots in the nucleus and a small number in the cytoplasm. We previously demonstrated that MTA1 distributed dynamically and periodically during the cell cycle. Here, through fluorescent colocalization, we found that MTA1 and Ku proteins colocalized well in the nucleus at interphase and moved synchronously from prophase to anaphase. Interestingly, at the time of telophase, when MTA1 was reported to re-enter the nucleus, they were separated and moved non-synchronously. Moreover, using in situ PLA, we visualized that the interaction occurred at both interphase and mitosis. At interphase, they interacted mainly in the nucleus, but during mitosis, they interact at the periphery of chromosomes. We also showed that MTA1 correlated well with Ku in both the cancerous and normal tissues, and that they cooperated in UV-induced DNA damage response. Collectively, our data uncover a specific interaction between MTA1 and Ku complex at both the nucleus and cytoplasm, and across the whole cell cycle. We therefore propose a potential functional crosstalk between NuRD and Ku complexes, the two most fundamental function units in cells, via physical interaction.
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影响因子：
8
作者：
Marzook, H.;Deivendran, S.;Pillai, M. R.
通讯作者：
Pillai, M. R.
影响因子：
3.9
作者：
Abdelbaqi K;Di Paola D;Rampakakis E;Zannis-Hadjopoulos M
通讯作者：
Zannis-Hadjopoulos M
DOI：
10.4161/cc.9.11.11735
发表时间：
2010-06-01
期刊：
Cell cycle (Georgetown, Tex.)
影响因子：
--
作者：
Li DQ;Kumar R
通讯作者：
Kumar R
DOI：
10.1073/pnas.0705878104
发表时间：
2007-08-07
影响因子：
11.1
作者：
Manavathi, Bramanandam;Peng, Shaohua;Kumar, Rakesh
通讯作者：
Kumar, Rakesh
影响因子：
8
作者：
Pucci, S;Mazzarelli, P;Fazio, VM
通讯作者：
Fazio, VM