Blinkowa A, Haldenwang W G, Ramsey J A, Henson J M, Mullin D A, Walker J R
J Bacteriol. 1983 Jan;153(1):66-75. doi: 10.1128/jb.153.1.66-75.1983.
Suppressors of a temperature-sensitive dnaZ polymerization mutant of Escherichia coli have been identified by selecting temperature-insensitive revertants. Those suppressed strains which concomitantly became cold sensitive were chosen for further study. Intragenic suppressor mutations, which caused cold-sensitive defects in DNA polymerization, were located in dnaZ by transduction with lambda dnaZ+ phages. Extragenic suppressor mutations were mapped within the initiation gene dnaA. These suppressor-containing strains were defective in initiation at low temperature as determined by measurements of DNA synthesis in vivo and in toluene-treated cells. The occurrence of suppressor mutations of dnaZ(Ts) within the dnaA gene is considered evidence that the dnaA and dnaZ products interact in vivo. A second indication of a dnaA-dnaZ protein-protein interaction was provided by the observation that the introduction of additional copies of the dnaZ+ gene into a strain carrying the dnaA suppressor mutation was lethal [whether the strain was dnaZ+ or dnaZ(Ts)].
通过筛选对温度不敏感的回复突变体,已鉴定出大肠杆菌温度敏感型dnaZ聚合突变体的抑制子。选择那些同时变得对冷敏感的抑制菌株进行进一步研究。通过用λ dnaZ⁺噬菌体进行转导,将导致DNA聚合出现冷敏感缺陷的基因内抑制突变定位在dnaZ中。基因外抑制突变定位在起始基因dnaA内。通过体内和经甲苯处理的细胞中的DNA合成测量确定,这些含有抑制子的菌株在低温下起始存在缺陷。dnaA基因内出现dnaZ(Ts)的抑制突变被认为是dnaA和dnaZ产物在体内相互作用的证据。dnaA-dnaZ蛋白质-蛋白质相互作用的第二个迹象是观察到,将额外拷贝的dnaZ⁺基因引入携带dnaA抑制突变的菌株中是致死的[无论该菌株是dnaZ⁺还是dnaZ(Ts)]。