Sakakibara Y
Department of Chemistry, National Institute of Health, Tokyo, Japan.
J Bacteriol. 1988 Feb;170(2):972-9. doi: 10.1128/jb.170.2.972-979.1988.
A newly isolated dnaK mutant of Escherichia coli, which contains the mutation dnaK111, has been found to be conditionally defective in initiation of DNA replication. Mutant cells that were transferred to high temperature exhibited residual DNA synthesis before the synthesis stopped completely. Analysis of the DNA synthesized at high temperature by hybridization with probe DNAs for detection of DNA replicated in the origin (oriC) and terminal (terC) regions has revealed that this mutant is unable to initiate a new round of DNA replication at high temperature after termination of the round in progress. The cells exposed to high temperature were subsequently capable of initiating DNA replication at low temperature in a synchronous manner. DNA synthesis of this mutant became temperature resistant upon inactivation of the rnh gene, similar to that of dnaA mutants, although cell growth of the dnaK mutant with the inactive rnh gene remained temperature sensitive. The dnaK mutation prevented DNA synthesis of lambda bacteriophage at high temperature even in the absence of the rnh gene function.
新分离出的含有dnaK111突变的大肠杆菌dnaK突变体,被发现其在DNA复制起始方面存在条件性缺陷。转移至高温环境的突变细胞在DNA合成完全停止前表现出残余的DNA合成。通过与用于检测在起始区域(oriC)和末端区域(terC)复制的DNA的探针DNA杂交,对高温下合成的DNA进行分析,结果显示该突变体在进行中的一轮DNA复制终止后,无法在高温下启动新一轮的DNA复制。暴露于高温的细胞随后能够在低温下以同步方式启动DNA复制。与dnaA突变体类似,当rnh基因失活时,该突变体的DNA合成变得对温度具有抗性,尽管rnh基因失活的dnaK突变体的细胞生长仍对温度敏感。即使在不存在rnh基因功能的情况下,dnaK突变也会在高温下阻止λ噬菌体的DNA合成。