Small molecule inhibitor of the RPA70 N-terminal protein interaction domain discovered using in silico and in vitro methods.
Small molecule inhibitor of the RPA70 N-terminal protein interaction domain discovered using in silico and in vitro methods.
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DOI：
10.1016/j.bmc.2011.03.012
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发表时间：
2011-04-15
影响因子：
3.5
通讯作者：
Oakley, Gregory G.
中科院分区：
文献类型：
--
作者：
Glanzer, Jason G.;Liu, Shengqin;Oakley, Gregory G.
药物对DNA损伤反应和DNA修复的抑制可以提高传统化疗药物的治疗指数。复制蛋白A(RPA)是真核生物中主要的单链DNA结合蛋白，在DNA复制、DNA修复、DNA重组和DNA损伤反应信号转导中起重要作用。通过对1,500个化合物的高通量筛选，我们已经确定了一种小分子抑制剂15-carboxy-13-isopropylatis-13-ene-17，18-dioic酸(NSC15520)，它可以抑制RPAN-末端DNA结合域(DBD)与Rad9-GST和P53-GST融合蛋白的结合，这些相互作用对于强大的DNA损伤信号是必不可少的。NSC15520竞争性地抑制P53-GST多肽与10μM的结合。NSC15520还抑制双链寡核苷酸的螺旋失稳，这种活性依赖于RPA70的N-末端结构域。NSC15520不抑制RPA与单链寡核苷酸的结合，提示该抑制剂的作用是针对RPA的N端DBD的，而不是与单链DNA结合所必需的DBD结合。计算机模拟表明NSC15520和Rad9之间直接竞争RPA上的同一结合表面。RPA N-末端的蛋白质-蛋白质相互作用的抑制剂被预测为与DNA损伤剂和DNA修复抑制剂具有协同作用。NSC15520等新型化合物具有作为化学增敏剂的潜力。
The pharmacological suppression of the DNA damage response and DNA repair can increase the therapeutic indices of conventional chemotherapeutics. Replication Protein A (RPA), the major single-stranded DNA binding protein in eukaryotes, is required for DNA replication, DNA repair, DNA recombination, and DNA damage response signaling. Through the use of high-throughput screening of 1500 compounds, we have identified a small molecule inhibitor, 15-carboxy-13-isopropylatis-13-ene-17,18-dioic acid (NSC15520), that inhibited both the binding of Rad9–GST and p53–GST fusion proteins to the RPA N-terminal DNA binding domain (DBD), interactions that are essential for robust DNA damage signaling. NSC15520 competitively inhibited the binding of p53-GST peptide with an IC50 of 10 μM. NSC15520 also inhibited helix destabilization of a duplex DNA (dsDNA) oligonucleotide, an activity dependent on the N-terminal domain of RPA70. NSC15520 did not inhibit RPA from binding single-stranded oligonucleotides, suggesting that the action of this inhibitor is specific for the N-terminal DBD of RPA, and does not bind to DBDs essential for single-strand DNA binding. Computer modeling implicates direct competition between NSC15520 and Rad9 for the same binding surface on RPA. Inhibitors of protein–protein interactions within the N-terminus of RPA are predicted to act synergistically with DNA damaging agents and inhibitors of DNA repair. Novel compounds such as NSC15520 have the potential to serve as chemosensitizing agents.
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影响因子：
64.8
作者：
Bartkova, J;Horejsi, Z;Bartek, J
通讯作者：
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影响因子：
4.8
作者：
Bochkareva, E;Frappier, L;Bochkarev, A
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作者：
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通讯作者：
Foulkes, William D.
影响因子：
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通讯作者：
Shirahige, Katsuhiko
影响因子：
64.8
作者：
Bochkarev, A;Pfuetzner, RA;Frappier, L
通讯作者：
Frappier, L