Human NTH1 physically interacts with p53 and proliferating cell nuclear antigen
Human NTH1 physically interacts with p53 and proliferating cell nuclear antigen
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DOI：
10.1016/j.bbrc.2004.06.136
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发表时间：
2004-08-11
影响因子：
3.1
通讯作者：
Matsunaga, T
中科院分区：
文献类型：
--
作者：
Oyama, M;Wakasugi, M;Matsunaga, T
胸腺嘧啶乙二醇（Tg）是电离辐射和其他氧化应激引起的主要DNA氧化损伤之一。人类NTH1是一种双功能酶，具有DNA糖基化酶和AP裂解酶的活性，并作为碱基切除修复（BER）的第一步去除Tg。我们通过下拉实验寻找与NTH1相互作用的因子，发现GST-NTH1融合蛋白沉淀增殖细胞核抗原（PCNA）和p53以及从人无细胞提取物中提取的XPG。GST-NTH1还与重组flag标记的XPG、PCNA和（His）(6)标记的p53蛋白结合，表明这些蛋白之间存在直接的蛋白-蛋白相互作用。此外，His-p53和FLAG-XPG刺激GST-NTH1或NTH1的Tg DNA糖基酶/AP裂解酶活性，而非PCNA。这些结果提供了对BER反应的积极调节的见解，也表明Tg的BER与其他细胞机制之间可能存在联系。(C) 2004爱思唯尔公司版权所有。
Thymine glycol (Tg) is one of predominant oxidative DNA lesions caused by ionizing radiation and other oxidative stresses. Human NTH1 is a bifunctional enzyme with DNA glycosylase and AP lyase activities and removes Tg as the first step of base excision repair (BER). We have searched for the factors interacting with NTH1 by using a pull-down assay and found that GST-NTH1 fusion protein precipitates proliferating cell nuclear antigen (PCNA) and p53 as well as XPG from human cell-free extracts. GST-NTH1 also bound to recombinant FLAG-tagged XPG, PCNA, and (His)(6)-tagged p53 proteins, indicating direct protein-protein interaction between those proteins. Furthermore, His-p53 and FLAG-XPG, but not PCNA, stimulated the Tg DNA glycosylase/AP lyase activity of GST-NTH1 or NTH1. These results provide an insight into the positive regulation of BER reaction and also suggest a possible linkage between BER of Tg and other cellular mechanisms. (C) 2004 Elsevier Inc. All rights reserved.