Control of initiation of F plasmid replication in Escherichia coli.

控制大肠杆菌中 F 质粒复制的起始。

• 批准号: 63580205
• 负责人: WADA Chieko
• 金额: $ 1.41万
• 依托单位: KYOTO UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (C)
• 财政年份: 1988
• 资助国家: 日本
• 起止时间: 1988 至 1990
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-63580205/
• 关键词: Replication initiation Of mini-F DNA; Transcription of repE gene; sigma 32 factor; Dnak protein; DnaJ protein; GrpE protein; Copy mutant of mini-F; ミニFコピ-変異; 大腸菌Fプラスミド; RepE蛋白質; σ^<32>RNAポリメラ-ゼ; FrepEプロモ-タ-; PlrepAプロモ-タ-; Rtsl repAプロモ-タ-; 太腸

Replication of F, mini-F and related plasmids ( mini-Rts1, mini-P1 ) is stringently controlled to maintain their copy number of 1 to 2 per host chromosome. It is thought that the plasmid copy number is regulated primarily by the amount or activity of the specific initiator protein essential for replication of the respective plasmid. We have found that the mini-F repE gene encoding the initiator protein is transcribed by RNA polymerase containing delta^<32> and that mini-F can not replicate in DELTA rpoH cells lacking delta^<32> protein. A subset of E.coli heat shock protein DnaK, DnaJ and GrpE are known to be required for lambda phage DNA replication and for growth of E.coli cells probably at all temperature. We have shown that these heat shock proteins are also required for replication of mini-F plasmid. When excess amounts of the replication initiator protein (RepE) were provided by means of a multicopy plasmid carrying repE,mini-F can replicate in the condition without DnaK, J, GrpE protein. Furthermore, we have isolated and characterized mini-F mutants able to replicate in the absence of delta^<32>. These mini-F plasmid produced altered initiator protein and exhibited a very high copy and were able to replicate in strains deficient in any of the above heat shock proteins. These results indicate that the subset of heat shock proteins play essential roles that help the functioning of the RepE initiator protein in mini-F DNA replication. We found that mini-Rts1 and mini-P1 like mini-F cannot replicate in the rpoH strain. The rep genes of both these plasmids like the mini-F were found to be transcribed by RNA polymerase delta^<32> as well as by delta^<70>. The replication of mini-Rts1 and mini-P1 like mini-F also depends on DnaK,DnaJ and GrpE protein.

严格控制F，Mini-F和相关质粒（Mini-RTS1，Mini-P1）的复制，以维持其每个宿主染色体1至2的拷贝数。据认为，质粒拷贝数主要由特定引发剂蛋白的数量或活性调节，对于复制相应的质粒必不可少。我们已经发现，编码引发剂蛋白的迷你-F基因被含有Delta^<32>的RNA聚合酶转录，并且在缺乏delta^<32>蛋白质的Delta RPOH细胞中，Mini-F无法复制。已知lambda噬菌体DNA复制和大肠杆菌细胞的生长可能在所有温度下可能是必需的。我们已经表明，这些热休克蛋白也需要复制Mini-F质粒。当通过多拷贝质粒复制提供过量的复制引发剂蛋白（REPE）时，Mini-F可以在没有DNAK，J，GRPE蛋白的条件下复制。此外，在没有delta^<32>的情况下，我们已经孤立并表征了能够复制的迷你F突变体。这些微型-F质粒产生了改变的引发剂蛋白质，并表现出很高的拷贝，并且能够在上述任何热休克蛋白中复制不足的菌株。这些结果表明，热休克蛋白的子集在Mini-F DNA复制中起作用，有助于Repe引发剂蛋白的功能。我们发现Mini-RTS1和Mini-P1像Mini-F一样无法在RPOH菌株中复制。发现这两种质粒（例如Mini-F）的代基因被RNA聚合酶delta^<32>以及Delta^<70>转录。 Mini-RTS1和Mini-P1的复制也取决于DNAK，DNAJ和GRPE蛋白。

项目成果

和田千恵子: "FプラスミドのDNA複製開始の調節" 生物物理. 28. 250-254 (1988)

Chieko Wada：“F 质粒 DNA 复制起始的调节”生物物理学 28. 250-254 (1988)。

Y.Kawasaki,C.Wada,M.Ishiai,and T.Yura: "Mechanism of DNA replication inhibition of F and related plasmids in a rpoH mutant." Japanese Journal of Genetics. 64-6. 493 (1989)

Y.Kawasaki、C.Wada、M.Ishiai 和 T.Yura：“rpoH 突变体中 F 和相关质粒的 DNA 复制抑制机制”。

T.Katayama, Y.Murakami, C.Wada, H.Ohmori, T.Yura, and T.Nagata: "Genetic suppression of a dnaG mutation in Escherichia coli" J. Bact. 171 1485-1491, 1989.

T.Katayama、Y.Murakami、C.Wada、H.Ohmori、T.Yura 和 T.Nagata：“大肠杆菌 dnaG 突变的基因抑制”J. Bact。

C.Wada: "Control of initiation of F plasmid replication" Seibutu Buturi, 28 250-254, 1988.

C.Wada：“F 质粒复制起始的控制”Seibutu Buturi，28 250-254，1988。

C.Wada;T.Yura: Annual report of the institute for virus research Kyoto University. 31. (1988)

C.Wada；T.Yura：京都大学病毒研究所年度报告。