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[TorR蛋白对大肠杆菌DNA复制起始的影响]

[The effects of TorR protein on initiation of DNA replication in Escherichia coli].

作者信息

Yao Yuan, Qiao Jia-Xin, Li Jing, Li Hui

机构信息

School of Life Sciences, Inner Mongolia University, Hohhot 010021, China.

出版信息

Yi Chuan. 2015 Mar;37(3):302-308. doi: 10.16288/j.yczz.14-291.

Abstract

The two-component systems, which could sense and respond to environmental changes, widely exist in bacteria as a signal transduction pathway. The bacterial CckA/CtrA, ArcA/ArcB and PhoP/PhoQ two-component systems are associated with initiation of DNA replication and cell division, however, the effects of the TorS/TorR system on cell cycle and DNA replication remains unknown. The TorS/TorR system in Escherichia coli can sense changes in trimethylamine oxide (TMAO) concentration around the cells. However, it is unknown if it also affects initiation of DNA replication. We detected DNA replication patterns in ΔtorS and ΔtorR mutant strains by flow cytometry. We found that the average number of replication origins (oriCs) per cell and doubling time in ΔtorS mutants were the same while the average number of oriCs in ΔtorR mutants was increased compared with that in wild-type cells. These results indicated that absence of TorR led to an earlier initiation of DNA replication than that in wild-type cells. Strangely, neither overexpression of TorR nor co-expression of TorR and TorS could restore ΔtorR mutant phenotype to the wild type. However, overexpression of SufD in both wild type and ΔtorR mutants promoted initiation of DNA replication, while mutation of SufD delayed it in ΔtorR mutants. Thus, TorR may affect initiation of DNA replication indirectly through regulating gene expression of sufD.

摘要

双组分系统能够感知并响应环境变化,作为一种信号转导途径广泛存在于细菌中。细菌的CckA/CtrA、ArcA/ArcB和PhoP/PhoQ双组分系统与DNA复制起始及细胞分裂相关,然而,TorS/TorR系统对细胞周期和DNA复制的影响尚不清楚。大肠杆菌中的TorS/TorR系统能够感知细胞周围氧化三甲胺(TMAO)浓度的变化。然而,其是否也影响DNA复制起始尚不清楚。我们通过流式细胞术检测了ΔtorS和ΔtorR突变株中的DNA复制模式。我们发现,ΔtorS突变株中每个细胞的复制起点(oriC)平均数量和倍增时间与野生型相同,而ΔtorR突变株中的oriC平均数量相较于野生型细胞增加。这些结果表明,缺失TorR导致DNA复制起始比野生型细胞更早。奇怪的是,TorR的过表达以及TorR和TorS的共表达均不能使ΔtorR突变体表型恢复为野生型。然而,野生型和ΔtorR突变株中SufD的过表达均促进了DNA复制起始,而SufD突变则在ΔtorR突变株中延迟了DNA复制起始。因此,TorR可能通过调节sufD的基因表达间接影响DNA复制起始。

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