Andrup L, Atlung T, Ogasawara N, Yoshikawa H, Hansen F G
Department of Microbiology, Technical University of Denmark, Lyngby.
J Bacteriol. 1988 Mar;170(3):1333-8. doi: 10.1128/jb.170.3.1333-1338.1988.
Plasmids carrying the intact Bacillus subtilis dnaA-like gene and two reciprocal hybrids between the B. subtilis and Escherichia coli dnaA genes were constructed. None of the plasmids could transform wild-type E. coli cells unless the cells contained surplus E. coli DnaA protein (DnaAEc). A dnaA (Ts) strain integratively suppressed by the plasmid R1 origin could be transformed by plasmids carrying either the B. subtilis gene (dnaABs) or a hybrid gene containing the amino terminus of the E. coli gene and the carboxyl terminus of the B. subtilis gene (dnaAEc/Bs). In cells with surplus E. coli DnaA protein, expression of the E. coli dnaA gene was derepressed by the B. subtilis DnaA protein and by the hybrid DnaAEc/Bs protein, whereas it was strongly repressed by the reciprocal hybrid protein DnaABs/Ec. The plasmids carrying the different dnaA genes probably all interfere with initiation of chromosome replication in E. coli by decreasing the E. coli DnaA protein concentration to a limiting level. The DnaABs and the DnaAEc/Bs proteins effect this decrease possibly by forming inactive oligomeric proteins, while the DnaABs/Ec protein may decrease dnaAEc gene expression.
构建了携带完整枯草芽孢杆菌dnaA样基因以及枯草芽孢杆菌和大肠杆菌dnaA基因之间两个相互杂交基因的质粒。除非细胞含有过量的大肠杆菌DnaA蛋白(DnaAEc),否则这些质粒均无法转化野生型大肠杆菌细胞。一个被质粒R1复制起点整合抑制的dnaA(Ts)菌株可被携带枯草芽孢杆菌基因(dnaABs)或包含大肠杆菌基因氨基末端和枯草芽孢杆菌基因羧基末端的杂交基因(dnaAEc/Bs)的质粒转化。在含有过量大肠杆菌DnaA蛋白的细胞中,大肠杆菌dnaA基因的表达被枯草芽孢杆菌DnaA蛋白和杂交DnaAEc/Bs蛋白去阻遏,而被反向杂交蛋白DnaABs/Ec强烈阻遏。携带不同dnaA基因的质粒可能都通过将大肠杆菌DnaA蛋白浓度降低到限制水平来干扰大肠杆菌染色体复制的起始。DnaABs和DnaAEc/Bs蛋白可能通过形成无活性的寡聚蛋白来实现这种降低,而DnaABs/Ec蛋白可能会降低dnaAEc基因的表达。