Screening of inhibitors of DNA replication by using the oriC plasmid replication system of Escherichia coli

Dna A蛋白是大肠杆菌中染色体DNA复制的启动子。本研究的目的是通过检测三磷酸腺苷与DNAA蛋白结合的抑制作用，发现在大肠杆菌中的DNA复制抑制物。由首席研究员小组进行的初步研究表明，一种蓝色的衍生物可以抑制DNAA蛋白的ATP结合活性。在这项研究中，我们合成了一些靛蓝衍生物，并研究了它们的结构与抑制DNAA蛋白的ATP结合的关系。我们证明了烷基残基是活性所必需的。结果，我们合成了一种在7微米时显示50%抑制作用的化合物。我们进一步证明了该化合物在ORIC质粒DNA复制系统中特异性地抑制了DNA合成的启动。研究结果于1996年发表在《生物化学》杂志上。

DnaA protein is the initiator of chromosomal DNA replication in Escherichia coli. The purpose of this study is a discovery of inhibitors of DNA replication in Escherichia coli by measurement of the inhibition of the ATP-binding to DnaA protein. Preliminary study by the group of the head investigator has shown that a derivative of indigo inhibits the ATP-binding activity of DnaA protein. In this study, we synthesized a number of indigo derivatives, and examined the relationship between their structures and inhibition of the ATP-binding to DnaA protein. We showed that alkyl residue is necessary for the activity. As a result, we could synthesize a compound which showed 50% inhibition at the concentration of 7muM.We further demonstrated that the compound specifically inhibited the initiation of DNA synthesis in an oriC plasmid DNA replication system. The results were published in J.Biol.Chem.in 1996.