Inhibitory Effects of Mismatch Binding Molecules on the Repair Reaction of Uracil-Containing DNA
Inhibitory Effects of Mismatch Binding Molecules on the Repair Reaction of Uracil-Containing DNA
复制标题
错配结合分子对含尿嘧啶DNA修复反应的抑制作用
DOI：
10.1021/acs.biochem.2c00344
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发表时间：
2022
期刊：
影响因子：
2.9
通讯作者：
Nakatani Kazuhiko
中科院分区：
文献类型：
--
作者：
Ulhusna Anisa;Murata Asako;Nakatani Kazuhiko
转录过程中形成的稳定 R 环会诱导酶介导的胞嘧啶脱氨基作用，产生的 DNA 中的尿嘧啶会激活碱基切除修复 (BER) 途径。 BER 途径中涉及的 DNA 切割被认为是三核苷酸重复不稳定的可能原因之一。在这里，我们进行了一项体外测定，以研究 DNA 结合小分子萘啶氨基甲酸酯二聚体 (NCD) 对 BER 酶反应的影响。凝胶电泳迁移率变动分析 (EMSA) 和热熔解分析揭示了 NCD 与 5'-XGG-3'/5'-XGG-3' 三联体（X = C 或 U 或无嘌呤/无嘧啶位点）的结合，该三联体是 BER 酶底物的模拟物。在 NCD 存在下，这些底物与 hSMUG1 和 APE1 酶的反应产物的聚丙烯酰胺凝胶电泳 (PAGE) 显示，NCD 干扰 5'-XGG-3'/5'-XGG-3' 三联体中的修复反应。这些发现将扩大小分子在调节三核苷酸重复不稳定性方面的潜力。
The stable R-loop formed during transcription induces enzyme-mediated deamination of cytosine, and the uracil in the DNA produced activates the base excision repair (BER) pathway. DNA cleavage involved in the BER pathway is thought to be one of the possible causes of trinucleotide repeat instability. Here, we performed an in vitro assay to investigate the effect of a DNA-binding small molecule, naphthyridine carbamate dimer (NCD), on BER enzyme reactions. The gel electrophoretic mobility shift assay (EMSA) and thermal melting analysis revealed the binding of NCD to a 5′-XGG-3′/5′-XGG-3′ triad (X = C or U or apurinic/apyrimidinic site), which is a mimic of a BER enzyme substrate. Polyacrylamide gel electrophoresis (PAGE) of the reaction products of these substrates with hSMUG1 and APE1 enzymes in the presence of NCD showed that NCD interfered with the repair reaction in the 5′-XGG-3′/5′-XGG-3′ triad. These findings would broaden the potential of small molecules in modulating trinucleotide repeat instability.