一种新的膜应激反应,通过ClpP蛋白酶靶向dna启动物来阻断染色体复制。

IF 2.7 3区 生物学 Q3 MICROBIOLOGY
Alabi Gbolahan, Tong Li, Rishit Saxena, Karen Wolcott, Aamna Sohail, Ishika Ahmed, Dhruba K Chattoraj, Elliott Crooke, Rahul Saxena
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引用次数: 0

摘要

在大肠杆菌中,由于脂蛋白(Lpp)成熟中断而引起的膜应激会损害DNA复制并阻止细胞生长。Lpp成熟与DNA复制之间的关系尚不清楚。我们证明,在膜胁迫下,Rcs应激反应途径被激活,复制启动子dna丢失,这解释了复制阻滞。然而,作为Rcs通路的关键调节因子,Lon蛋白酶并不是dna丢失所必需的。我们进一步排除了细菌应激反应的主要介质之一(p)ppGpp的参与。另一方面,在ClpP蛋白酶基因缺失后,dna是稳定的,复制没有被抑制,也没有细胞生长停滞。在野生型细胞中,即使没有膜应激,dna的过表达也是致命的。在∆crp细胞中,过度起始受到抑制,dna的过表达能够克服生长停滞。先前发现的抗膜胁迫的∆fis细胞,经过胁迫诱导处理后dna稳定,复制正常。我们得出结论,dna的丢失足以解释膜应力下的生长停滞。这里描述的应激反应途径似乎是新颖的,因为它独立于Lon和(p)ppGpp,这两种物质与其他阻断DNA复制的应激反应有关。重要性:DNA复制胁迫可以阻断大肠杆菌的细胞分裂(SOS反应),这一发现引入了细胞周期中检查点控制的概念。在这里,我们描述了一种新的检查点控制,它在相反的方向上起作用:膜应力导致复制阻滞。我们展示了前体脂蛋白(pLpp)的积累如何阻止复制。pLpp积累引起反应,最终激活ClpP蛋白酶,通过靶向启动dna来阻止复制。dna是至关重要且高度保守的,对反应途径的详细了解可能为治疗细菌感染开辟新的途径。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
A novel membrane stress response that blocks chromosomal replication by targeting the DnaA initiator via the ClpP protease.

In Escherichia coli, membrane stress due to interrupted lipoprotein (Lpp) maturation impairs DNA replication and arrests cell growth. How Lpp maturation and DNA replication are connected remains unclear. We demonstrate that upon membrane stress, the Rcs stress-response pathway is activated, and the replication initiator DnaA is lost, which explains the replication block. However, Lon protease, a key regulator of the Rcs pathway, is not required for the DnaA loss. We further ruled out the involvement of (p)ppGpp, one of the major mediators of stress responses in bacteria. On the other hand, upon deletion of the ClpP protease gene, DnaA was stable, replication was not inhibited, and there was no cell-growth arrest. In wild-type cells, overexpression of DnaA was lethal even without the membrane stress apparently from hyperinitiation. In ∆crp cells, hyperinitiation was restrained, and overexpression of DnaA was able to overcome the growth arrest. ∆fis cells, which were earlier found resistant to the membrane stress, showed DnaA stability and normal replication upon stress-inducing treatments. We conclude that DnaA loss suffices to explain the growth arrest upon the membrane stress. The stress-response pathway described here appears novel because of its independence from Lon and (p)ppGpp, which have been implicated in other stress responses that block DNA replication.

Importance: The observation that DNA replication stress can block cell division in E. coli (SOS response) introduced the concept of checkpoint control in the cell cycle. Here, we describe a novel checkpoint control that functions in the opposite direction: membrane stress causing replication block. We show how the accumulation of precursor lipoprotein (pLpp) could block replication. The pLpp accumulation causes a response culminating in activating the ClpP protease that blocks replication by targeting the initiator DnaA. DnaA being vital and highly conserved, a detailed understanding of the response pathway is likely to open new avenues to treat bacterial infection.

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来源期刊
Journal of Bacteriology
Journal of Bacteriology 生物-微生物学
CiteScore
6.10
自引率
9.40%
发文量
324
审稿时长
1.3 months
期刊介绍: The Journal of Bacteriology (JB) publishes research articles that probe fundamental processes in bacteria, archaea and their viruses, and the molecular mechanisms by which they interact with each other and with their hosts and their environments.
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