Identification of a small-molecule inhibitor that selectively blocks DNA-binding by Trypanosoma brucei replication protein A1.
Identification of a small-molecule inhibitor that selectively blocks DNA-binding by Trypanosoma brucei replication protein A1.
复制标题
DOI：
10.1038/s41467-023-39839-x
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发表时间：
2023-07-20
影响因子：
16.6
通讯作者：
Kim, Hee-Sook
中科院分区：
文献类型：
--
作者：
Mukherjee, Aditi;Hossain, Zakir;Erben, Esteban;Ma, Shuai;Choi, Jun Yong;Kim, Hee-Sook
复制蛋白A （RPA）是由RPA1、2和3亚基组成的广泛保守的复合体。RPA在DNA复制和修复过程中保护暴露的单链DNA （ssDNA）。通过结构建模，我们发现了一种抑制剂JC-229，它可以靶向布鲁氏锥虫（非洲锥虫病的致病寄生虫）中的RPA1。该抑制剂对布鲁氏t细胞有高毒性，对人体细胞有轻度毒性。JC-229处理模拟了TbRPA1缺失的影响，包括DNA复制抑制和DNA损伤积累。体外ssdna结合实验表明，JC-229抑制TbRPA1的活性，但不抑制人类同源物的活性。事实上，尽管JC-229与T. cruzi和利什曼原虫RPA1的序列高度同源，但它只影响TbRPA1的ssdna结合活性。定点突变证实，TbRPA1中的dna结合域A （DBD-A）含有一个JC-229结合口袋。残基丝氨酸105决定TbRPA1的特异性结合和抑制，但不影响克氏T.和利什曼原虫RPA1。我们的数据为开发和测试用于治疗非洲锥虫病的高度特异性抑制剂提供了一条途径。作者鉴定了一种小分子抑制剂，靶向布鲁氏T.复制蛋白A1的ssdna结合功能，而不影响宿主的同源性。他们确认了TbRPA1中的关键残基与化学探针的物种特异性选择性有关。
Replication Protein A (RPA) is a broadly conserved complex comprised of the RPA1, 2 and 3 subunits. RPA protects the exposed single-stranded DNA (ssDNA) during DNA replication and repair. Using structural modeling, we discover an inhibitor, JC-229, that targets RPA1 in Trypanosoma brucei, the causative parasite of African trypanosomiasis. The inhibitor is highly toxic to T. brucei cells, while mildly toxic to human cells. JC-229 treatment mimics the effects of TbRPA1 depletion, including DNA replication inhibition and DNA damage accumulation. In-vitro ssDNA-binding assays demonstrate that JC-229 inhibits the activity of TbRPA1, but not the human ortholog. Indeed, despite the high sequence identity with T. cruzi and Leishmania RPA1, JC-229 only impacts the ssDNA-binding activity of TbRPA1. Site-directed mutagenesis confirms that the DNA-Binding Domain A (DBD-A) in TbRPA1 contains a JC-229 binding pocket. Residue Serine 105 determines specific binding and inhibition of TbRPA1 but not T. cruzi and Leishmania RPA1. Our data suggest a path toward developing and testing highly specific inhibitors for the treatment of African trypanosomiasis. The authors identify a small molecule inhibitor targeting the ssDNA-binding function of T. brucei Replication Protein A1 without affecting the host ortholog. They confirm the key residue in TbRPA1 involved in the species-specific selectivity of the chemical probe.
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影响因子：
2.6
作者：
Cestari I;Stuart K
通讯作者：
Stuart K
影响因子：
2.9
作者：
Souza-Fagundes EM;Frank AO;Feldkamp MD;Dorset DC;Chazin WJ;Rossanese OW;Olejniczak ET;Fesik SW
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3.5
作者：
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通讯作者：
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DOI：
10.1016/j.bbagen.2020.129607
发表时间：
2020-07-01
影响因子：
3
作者：
Fernandes, Carlos A. H.;Morea, Edna Gicela O.;Cano, Maria Isabel Nogueira
通讯作者：
Cano, Maria Isabel Nogueira
影响因子：
4.8
作者：
Carter M;Gomez S;Gritz S;Larson S;Silva-Herzog E;Kim HS;Schulz D;Hovel-Miner G
通讯作者：
Hovel-Miner G