Molecular Design of Inhibitors of in Vitro oriC DNA Replication Based on the Potential to Block the ATP Binding of DnaA Protein*
Molecular Design of Inhibitors of in Vitro oriC DNA Replication Based on the Potential to Block the ATP Binding of DnaA Protein*
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基于阻断 DnaA 蛋白 ATP 结合潜力的体外 oriC DNA 复制抑制剂的分子设计*
DOI：
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发表时间：
1996
影响因子：
4.8
通讯作者：
K. Sekimizu
中科院分区：
文献类型：
--
作者：
T. Mizushima;S. Sasaki;Hiroko Ohishi;M. Kobayashi;T. Katayama;T. Miki;M. Maeda;K. Sekimizu
Dna A蛋白是大肠杆菌中染色体DNA复制的起始因子，通过与ATP结合而被激活。我们早些时候报道了3-乙酰氧基-2，2‘-双-1H-吲哚抑制DNAA蛋白与ATP的结合(Sasaki，S.，Mizushima，T.，Hashimoto，T.，Maeda，M.和Sekimizu，K.(1994)Bioorg)。地中海医院。化学。让我们来吧。4、1771-1774)。本研究合成了3-乙酰氧基-2，2‘-双-1H-吲哚类化合物，并研究了它们对DNAA蛋白与ATP结合的抑制作用。脂肪链的延长导致低浓度下ATP与DNAA蛋白的结合受到抑制。其中3-[N-(11-carboxyundecyl)]carbamoylmethoxy-2，2′-bi-1H-indol(结构7(3-CUCM-BI))的抑制作用最强，其IC50值为7μM，其抑制方式为竞争性。我们进一步证明了结构7(3-CUCM-BI)在由纯化的蛋白质重组的系统中抑制了ORIC质粒的DNA复制。这种抑制作用是针对DNA复制的启动，而不是针对DNA的延伸。这种抑制作用可通过DNAA蛋白与三磷酸腺苷预先孵育来克服。此外，结构7(3-CUCM-BI)对ABC系统的DNA合成几乎没有抑制作用。我们认为结构7(3-CUCM-BI)通过抑制ATP与DNAA蛋白的结合而在体外ORIC DNA复制中发挥作用。
DnaA protein, the initiation factor for chromosomal DNA replication in Escherichia coli, is activated by binding to ATP. We earlier reported that 3-acetoxy-2,2′-bi-1H-indol inhibited the ATP binding to DnaA protein (Sasaki, S., Mizushima, T., Hashimoto, T., Maeda, M., and Sekimizu, K. (1994) Bioorg. Med. Chem. Lett. 4, 1771-1774). In the present study, derivatives of 3-acetoxy-2,2′-bi-1H-indol with different lengths of aliphatic chains at the 3-O position were synthesized, and their potential to inhibit the ATP binding to DnaA protein was examined. Elongation of the aliphatic chain resulted in inhibition of the ATP binding to DnaA protein at lower concentrations. Among the derivatives, 3-[N-(11-carboxyundecyl)]carbamoylmethoxy-2,2′-bi-1H-indol (structure 7 (3-CUCM-BI)) exhibited the most potent inhibition with an IC50 value of 7 μM. The mode of the inhibition was competitive. We further demonstrated that structure 7 (3-CUCM-BI) inhibited DNA replication of the oriC plasmid in a system reconstituted from purified proteins. This inhibition was specific for the initiation of DNA replication rather than for the elongation. The inhibition was overcome by preincubation of DnaA protein with ATP. Furthermore, structure 7 (3-CUCM-BI) showed little inhibition on DNA synthesis in the ABC primosome system. We propose that structure 7 (3-CUCM-BI) functions in the in vitro oriC DNA replication by inhibiting the ATP binding to DnaA protein.
膜附着激活 dnaA 蛋白，这是大肠杆菌中染色体复制的起始蛋白。
DOI：
10.1073/pnas.85.19.7202
发表时间：
1988
影响因子：
11.1
作者：
Yung,BY;Kornberg,A
通讯作者：
Kornberg,A