TOX4 and its binding partners recognize DNA adducts generated by platinum anticancer drugs
TOX4 and its binding partners recognize DNA adducts generated by platinum anticancer drugs
复制标题
DOI：
10.1016/j.abb.2010.12.021
复制
发表时间：
2011-03-15
影响因子：
3.9
通讯作者：
Breton, Jean
中科院分区：
文献类型：
--
作者：
du Puch, Christophe Bounaix Morand;Barbier, Ewa;Breton, Jean
铂剂是通常开出的破坏DNA的抗癌药物。诱导的损伤被广泛的蛋白质识别。这些都涉及细胞机制，如DNA修复、调节细胞毒性或染色质重塑。因此，它们构成了理解这些药物的药理学的关键因素。为了扩大我们对这个亚蛋白质组的了解，我们开发了一个与高通量蛋白质组工具相结合的配基捕鱼器。这种捕捉器是由附着在磁珠上的受损质粒制成的，并暴露在细胞核提取液中。保留的蛋白质用纳米高效液相色谱-串联质谱仪进行鉴定。这种方法使我们建立了一份38种蛋白质的清单，这些蛋白质与顺铂、奥沙利铂和赛特铂产生的DNA加合物相互作用。其中一些是已知的相互作用组成员，如高迁移率组蛋白1(HMGB1)或人类上游结合因子(HUBF)，但我们也成功地鉴定了一些意想不到的蛋白质，如TOX HMG盒家族成员4(TOX4)、磷酸酶1核靶向亚单位(PNUTS)和WD包含重复的蛋白82(WDR82)，这些都是最近发现的复合体的成员。随后，通过表面等离子体共振成像(SPRI)验证了TOX4与铂化DNA之间的相互作用。这些相互作用突显了细胞对化疗药物诱导的DNA损伤的新反应。(C)2010 Elsevier Inc.保留所有权利。
Platinating agents are commonly prescribed anticancer drugs damaging DNA. Induced lesions are recognized by a wide range of proteins. These are involved in cellular mechanisms such as DNA repair, mediation of cytotoxicity or chromatin remodeling. They therefore constitute crucial actors to understand pharmacology of these drugs. To expand our knowledge about this subproteome, we developed a ligand fishing trap coupled to high throughput proteomic tools. This trap is made of damaged plasmids attached to magnetic beads, and was exposed to cell nuclear extracts. Retained proteins were identified by nano-HPLC coupled to tandem mass spectrometry. This approach allowed us to establish a list of 38 proteins interacting with DNA adducts generated by cisplatin, oxaliplatin and satraplatin. Some of them were already known interactome members like high mobility group protein 1 (HMGB1) or the human upstream binding factor (hUBF), but we also succeeded in identifying unexpected proteins such as TOX HMG box family member 4 (TOX4), phosphatase 1 nuclear targeting subunit (PNUTS), and WD repeat-containing protein 82 (WDR82), members of a recently discovered complex. Interaction between TOX4 and platinated DNA was subsequently validated by surface plasmon resonance imaging (SPRi). These interactions highlight new cellular responses to DNA damage induced by chemotherapeutic agents. (C) 2010 Elsevier Inc. All rights reserved.