Mapping the Ku Interactome Using Proximity-Dependent Biotin Identification in Human Cells.,Journal of Proteome Research

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Mapping the Ku Interactome Using Proximity-Dependent Biotin Identification in Human Cells.
Journal of Proteome Research ( IF 3.8 ) Pub Date : 2019-01-14 , DOI: 10.1021/acs.jproteome.8b00771
Sanna Abbasi ₁ , Caroline Schild-Poulter ₁

Affiliation

The Ku heterodimer, composed of Ku70 and Ku80, is best characterized for its role in repairing double-stranded DNA breaks but is also known to participate in other regulatory processes. Despite our understanding of Ku protein interplay during DNA repair, the extent of Ku's protein interactions in other processes has never been fully determined. Using proximity-dependent biotin identification (BioID) and affinity purification coupled to mass spectrometry (AP-MS) with wild-type Ku70, we identified candidate proteins that interact with the Ku heterodimer in HEK293 cells, in the absence of exogenously induced DNA damage. BioID analysis identified approximately 250 nuclear proteins, appearing in at least two replicates, including known Ku-interacting factors such as MRE11A, WRN, and NCOA6. Meanwhile, AP-MS analysis identified approximately 50 candidate proteins. Of the novel protein interactors identified, many were involved in functions already suspected to involve Ku such as transcriptional regulation, DNA replication, and DNA repair, while several others suggest that Ku may be involved in additional functions such as RNA metabolism, chromatin-remodeling, and microtubule dynamics. Using a combination of BioID and AP-MS, this is the first report that comprehensively characterizes the Ku protein interaction landscape, revealing new cellular processes and protein complexes involving the Ku complex.

中文翻译：

在人体细胞中使用邻近依赖性生物素鉴定定位Ku相互作用组。

由Ku70和Ku80组成的Ku异二聚体，以其在修复双链DNA断裂中的作用而闻名，但也参与其他调控过程。尽管我们了解在DNA修复过程中Ku蛋白的相互作用，但尚未完全确定Ku在其他过程中蛋白相互作用的程度。使用邻近依赖性生物素鉴定（BioID）和亲和纯化与质谱（AP-MS）结合的野生型Ku70，我们确定了在没有外源诱导的DNA损伤的情况下与HEK293细胞中Ku异二聚体相互作用的候选蛋白。BioID分析鉴定出大约250种核蛋白，至少以两个重复出现，包括已知的Ku相互作用因子，例如MRE11A，WRN和NCOA6。同时，AP-MS分析鉴定出大约50种候选蛋白质。在鉴定出的新型蛋白质相互作用子中，许多参与了已经怀疑与Ku有关的功能，例如转录调控，DNA复制和DNA修复，而另一些则表明Ku可能参与了其他功能，如RNA代谢，染色质重塑，和微管动力学。这是使用BioID和AP-MS的组合，这是第一份全面描述Ku蛋白相互作用图景的报告，揭示了涉及Ku复合物的新细胞过程和蛋白复合物。而其他一些人则暗示Ku可能参与了其他功能，例如RNA代谢，染色质重塑和微管动力学。这是使用BioID和AP-MS的组合，这是第一份全面描述Ku蛋白相互作用图景的报告，揭示了涉及Ku复合物的新细胞过程和蛋白复合物。而其他一些人则暗示Ku可能参与了其他功能，例如RNA代谢，染色质重塑和微管动力学。这是使用BioID和AP-MS的组合，这是第一份全面描述Ku蛋白相互作用图景的报告，揭示了涉及Ku复合物的新细胞过程和蛋白复合物。

更新日期：2019-02-07

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