Evaluation of simultaneous binding of Chromomycin A3 to the multiple sites of DNA by the new restriction enzyme assay
Evaluation of simultaneous binding of Chromomycin A3 to the multiple sites of DNA by the new restriction enzyme assay
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DOI：
10.1016/j.bmcl.2018.04.013
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发表时间：
2018-06-01
影响因子：
2.7
通讯作者：
Sasaki, Shigeki
中科院分区：
文献类型：
--
作者：
Murase, Hirotaka;Noguchi, Tomoharu;Sasaki, Shigeki
色霉素A3（Chromomycin A3，CMA 3）是一种金油酸型抗肿瘤抗生素。CMA 3与二价阳离子如Mg 2+形成二聚体复合物，其强烈结合DNA的富含GC的序列以抑制DNA复制和转录。在这项研究中，通过测量限制性酶AccII和Fnu 4 HI分别对CGCG位点和5 '-GC向下箭头GGC位点的中心的水解保护，研究了CMA 3与含有多个富含GC的结合位点的DNA序列的结合特性。与标准DNA酶I足迹法相反，DNA底物被限制性酶完全水解，因此，在所有可切割位点处的DNA的完全保护表明CMA 3同时结合所有结合位点。限制性内切酶分析表明，CMA 3具有结合连续的CGCG位点和CGG重复序列的高倾向。(C)2018爱思唯尔有限公司版权所有。
Chromomycin A3 (CMA3) is an aureolic acid-type antitumor antibiotic. CMA3 forms dimeric complexes with divalent cations, such as Mg2+, which strongly binds to the GC rich sequence of DNA to inhibit DNA replication and transcription. In this study, the binding property of CMA3 to the DNA sequence containing multiple GC-rich binding sites was investigated by measuring the protection from hydrolysis by the restriction enzymes, AccII and Fnu4HI, for the center of the CGCG site and the 5'-GC down arrow GGC site, respectively. In contrast to the standard DNase I footprinting method, the DNA substrates are fully hydrolyzed by the restriction enzymes, therefore, the full protection of DNA at all the cleavable sites indicates that CMA3 simultaneously binds to all the binding sites. The restriction enzyme assay has suggested that CMA3 has a high tendency to bind the successive CGCG sites and the CGG repeat. (C) 2018 Elsevier Ltd. All rights reserved.