Isolation and detection of DNA-protein crosslinks in mammalian cells.
Isolation and detection of DNA-protein crosslinks in mammalian cells.
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DOI：
10.1093/nar/gkad1178
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发表时间：
2024-01-25
影响因子：
14.9
通讯作者：
中科院分区：
文献类型：
--
作者：
DNA-蛋白质交联（DPC）是毒性DNA损伤，其中蛋白质共价连接到DNA。如果不能迅速修复，DPC会产生干扰DNA复制、转录和DNA损伤修复的障碍，最终导致基因组不稳定。DPC的持续存在与过早衰老、癌症和神经变性有关。在哺乳动物细胞中，DPC的修复主要依赖于SPRTN和26 S蛋白酶体的蛋白水解活性，并辅以其他酶，包括TDP 1/2和MRN复合物，并且所涉及的许多活性是必需的，限制了遗传方法。多年来，哺乳动物细胞中DPC修复的研究受到缺乏标准化测定的阻碍，最值得注意的是可靠地定量与DNA共价结合的蛋白质或蛋白水解片段的测定。最近在该领域的兴趣已经刺激了DPC分析的几种生物化学方法的发展。在这里，我们批判性地分析了DPC隔离的最新技术以及每种技术的优缺点。我们的目标是帮助研究人员选择最适合他们的实验要求和问题的分离方法，并促进不同实验室使用不同方法的结果比较。
DNA–protein crosslinks (DPCs) are toxic DNA lesions wherein a protein is covalently attached to DNA. If not rapidly repaired, DPCs create obstacles that disturb DNA replication, transcription and DNA damage repair, ultimately leading to genome instability. The persistence of DPCs is associated with premature ageing, cancer and neurodegeneration. In mammalian cells, the repair of DPCs mainly relies on the proteolytic activities of SPRTN and the 26S proteasome, complemented by other enzymes including TDP1/2 and the MRN complex, and many of the activities involved are essential, restricting genetic approaches. For many years, the study of DPC repair in mammalian cells was hindered by the lack of standardised assays, most notably assays that reliably quantified the proteins or proteolytic fragments covalently bound to DNA. Recent interest in the field has spurred the development of several biochemical methods for DPC analysis. Here, we critically analyse the latest techniques for DPC isolation and the benefits and drawbacks of each. We aim to assist researchers in selecting the most suitable isolation method for their experimental requirements and questions, and to facilitate the comparison of results across different laboratories using different approaches.
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影响因子：
5.9
作者：
John, Tobias;Pires, Elisabete;Hester, Svenja S.;Salah, Eidarus;Hopkinson, Richard J.;Schofield, Christopher J.
通讯作者：
Schofield, Christopher J.
影响因子：
64.5
作者：
Dheekollu J;Wiedmer A;Ayyanathan K;Deakyne JS;Messick TE;Lieberman PM
通讯作者：
Lieberman PM
影响因子：
3.8
作者：
Fielden J;Ruggiano A;Popović M;Ramadan K
通讯作者：
Ramadan K
影响因子：
4.8
作者：
Barker, S;Weinfeld, M;Murray, D
通讯作者：
Murray, D
影响因子：
16.2
作者：
Aditi;Downing SM;Schreiner PA;Kwak YD;Li Y;Shaw TI;Russell HR;McKinnon PJ
通讯作者：
McKinnon PJ