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首页> 外文期刊>Journal of bacteriology >Incompatibility Protein IncC and Global Regulator KorB Interact in Active Partition of Promiscuous Plasmid RK2
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Incompatibility Protein IncC and Global Regulator KorB Interact in Active Partition of Promiscuous Plasmid RK2

机译：不相容蛋白IncC和全局调节剂KorB在混杂质粒RK2的活性分配中相互作用。

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Replication of the broad-host-range, IncPα plasmid RK2 requires two plasmid loci: trfA, the replication initiator gene, andoriV, the origin of replication. While these determinants are sufficient for replication in a wide variety of bacteria, they do not confer the stable maintenance of parental RK2 observed in its hosts. The product of the incC gene has been proposed to function in the stable maintenance of RK2 because of its relatedness to the ParA family of ATPases, some of which are known to be involved in the active partition of plasmid and chromosomal DNA. Here we show that IncC has the properties expected of a component of an active partition system. The smaller polypeptide product of incC (IncC2) exhibits a strong, replicon-independent incompatibility phenotype with RK2. This incompatibility phenotype requires the global transcriptional repressor, KorB, and the target for incC-mediated incompatibility is a KorB-binding site (OB). We found that KorB and IncC interact in vivo by using the yeast two-hybrid system and in vitro by using partially purified proteins. Elevated expression of the incC and korB genes individually has no obvious effect on Escherichia coli cell growth, but their simultaneous overexpression is toxic, indicating a possible interaction of IncC-KorB complexes with a vital host target. A region of RK2 bearing incC, korB, and multiple KorB-binding sites is able to stabilize an unstable, heterologous plasmid in anincC-dependent manner. Finally, elevated levels of IncC2 cause RK2 to aggregate, indicating a possible role for IncC in plasmid pairing. These findings demonstrate that IncC, KorB, and at least one KorB-binding site are components of an active partition system for the promiscuous plasmid RK2.
机译：复制宽宿主范围的IncPα质粒RK2需要两个质粒基因座：复制起始基因 trfA 和复制起点 oriV 。尽管这些决定因素足以在多种细菌中复制，但它们不能赋予宿主RK2亲本RK2稳定的维持能力。由于 incC 基因的产物与RKase的ParA家族相关，因此有人提出其可以在RK2的稳定维持中发挥作用，其中一些已知与质粒的活性分配有关。染色体DNA。在这里，我们显示IncC具有活动分区系统的组件所期望的属性。 incC （IncC2）的较小多肽产物与RK2表现出强的，不依赖复制子的不相容表型。这种不相容性表型需要全局转录阻遏物KorB，而 incC 介导的不相容性的靶标是KorB结合位点（O B ）。我们发现，KorB和IncC通过使用酵母双杂交系统在体内相互作用，而在体外通过使用部分纯化的蛋白相互作用。分别 incC 和 korB 基因的表达升高对大肠杆菌细胞的生长没有明显影响，但是它们同时过表达具有毒性，表明可能IncC-KorB复合物与重要宿主目标的相互作用。带有 incC ， korB 和多个KorB结合位点的RK2区域能够稳定依赖于 incC 的不稳定，异源质粒方式。最后，IncC2水平升高会导致RK2聚集，表明IncC在质粒配对中可能发挥作用。这些发现表明，IncC，KorB和至少一个KorB结合位点是混杂质粒RK2的活性分配系统的组分。

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