López-Villarejo Juan, Lobato-Márquez Damián, Díaz-Orejas Ramón
Department of Molecular Microbiology and Infection Biology, C/ Ramiro de Maéztu 9, Centro de Investigaciones Biológicas-CSIC, 28040 Madrid, Spain.
Toxins (Basel). 2015 Feb 5;7(2):478-92. doi: 10.3390/toxins7020478.
kis-kid, the auxiliary maintenance system of plasmid R1 and copB, the auxiliary copy number control gene of this plasmid, contribute to increase plasmid replication efficiency in cells with lower than average copy number. It is thought that Kis antitoxin levels decrease in these cells and that this acts as the switch that activates the Kid toxin; activated Kid toxin reduces copB-mRNA levels and this increases RepA levels that increases plasmid copy number. In support of this model we now report that: (i) the Kis antitoxin levels do decrease in cells containing a mini-R1 plasmid carrying a repA mutation that reduces plasmid copy number; (ii) kid-dependent replication rescue is abolished in cells in which the Kis antitoxin levels or the CopB levels are increased. Unexpectedly we found that this coordination significantly increases both the copy number of the repA mutant and of the wt mini-R1 plasmid. This indicates that the coordination between plasmid replication functions and kis-kid system contributes significantly to control plasmid R1 replication.
Kis-kid是质粒R1的辅助维持系统,也是该质粒的辅助拷贝数控制基因,有助于提高拷贝数低于平均水平的细胞中的质粒复制效率。据认为,这些细胞中Kis抗毒素水平降低,这充当激活Kid毒素的开关;被激活的Kid毒素降低copB-mRNA水平,进而增加RepA水平,从而增加质粒拷贝数。为支持该模型,我们现在报告:(i)在含有携带降低质粒拷贝数的repA突变的微型R1质粒的细胞中,Kis抗毒素水平确实降低;(ii)在Kis抗毒素水平或CopB水平增加的细胞中,kid依赖性复制拯救被消除。出乎意料的是,我们发现这种协调显著增加了repA突变体和野生型微型R1质粒的拷贝数。这表明质粒复制功能与kis-kid系统之间的协调对控制质粒R1复制有显著贡献。