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

利用大肠杆菌oriC质粒复制系统筛选DNA复制抑制剂

• 批准号: 06557130
• 负责人: SEKIMIZU Kazuhisa
• 金额: $ 6.53万
• 依托单位: KYUSHU UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (A)
• 财政年份: 1994
• 资助国家: 日本
• 起止时间: 1994 至 1996
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-06557130/
• 关键词: DnaA protein; DNA replication; chromosomal DNA; Escherichia coli.; ATP-binding; indigo; oriC plasmid; oriCプラスミド; DNA合成阻害剤; 脂質クラスター; ovic複製; oriC; DNAポリメラーゼ; 脂質; dnaA変異株

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.

DnaA 蛋白是大肠杆菌中染色体 DNA 复制的起始因子。本研究的目的是通过测量 ATP 与 DnaA 蛋白结合的抑制来发现大肠杆菌中 DNA 复制的抑制剂。首席研究员小组的初步研究表明，靛蓝衍生物会抑制 DnaA 蛋白的 ATP 结合活性。在本研究中，我们合成了多种靛蓝衍生物，并研究了它们的结构与抑制 ATP 与 DnaA 蛋白结合的关系。我们证明烷基残基对于该活性是必需的。结果，我们合成了一种在7μM浓度下表现出50％抑制作用的化合物。我们进一步证明该化合物特异性抑制oriC质粒DNA复制系统中DNA合成的起始。结果于1996年发表在J.Biol.Chem.上。

项目成果

T.Mizushima, J.Tanino, T.Miki, and K.Sekimizu: "Decrease in the linking number of plasmid DNA in dnaA mutants of Escherichia coli" Biochem.Biophys.Res.Commun.218. 137-141 (1996)

T.Mizushima、J.Tanino、T.Miki 和 K.Sekimizu：“大肠杆菌 dnaA 突变体中质粒 DNA 连接数量的减少”Biochem.Biophys.Res.Commun.218。

Tohru Mizushima,S.Sasaki,… & Kazuhisa Sekimizu: "Molecular Design of Inhibitors of in Vitro oriC DNA Replication Based on the Potential to Block the ATPB Binding of DnaA Protein" The Journal of Biological Chemistry. 271. 25178-25183 (1996)

Tohru Mizushima、S.Sasaki、… & Kazuhisa Sekimizu：“基于阻断 DnaA 蛋白 ATPB 结合潜力的体外 oriC DNA 复制抑制剂的分子设计”《生物化学杂志》271。25178-25183 (1996)。

T.Mizushima,J.Tamino,T.Miki & K.Sekimizu: "Decrease in the linking number of phasmid DNA in dnaA mutants of Escherihica coli" Biophys,Biochen.Res.Commun. (in press). (1996)

T.水岛、J.塔米诺、T.三木

T.Mizushima,A.Tomura,T.Shimpuku,T.Miki and K.Sekimizu: "Loss of flagellation in dneA mutants of Eschetichia coli." J. Bacteriol.176. 5544-5546 (1994)

T.Mizushima、A.Tomura、T.Shimpuku、T.Miki 和 K.Sekimizu：“大肠杆菌 dneA 突变体中鞭毛的丢失。”

S.Takeda,K.Kita,H.Miyazaki,S.Natori and K.Sekimizu: "Purification of low-molecular-weight GTP-binding proteins structurally related to MTG 33,amitochoudrial GTP-binding protein of Ehrlich ascites turncells" J.Biochem.(Tokyo). 118. 791-795 (1995)

S.Takeda、K.Kita、H.Miyazaki、S.Natori 和 K.Sekimizu：“与 Ehrlich 腹水转细胞的 MTG 33、amitochoudrial GTP 结合蛋白结构相关的低分子量 GTP 结合蛋白的纯化” J.