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新月柄杆菌 CdnL 是一种非必需的 RNA 聚合酶结合蛋白,其缺失会损害正常生长和 rRNA 转录。

Caulobacter crescentus CdnL is a non-essential RNA polymerase-binding protein whose depletion impairs normal growth and rRNA transcription.

机构信息

Departamento de Genética y Microbiología, Área de Genética (Unidad Asociada al IQFR-CSIC), Facultad de Biología, Universidad de Murcia, 30100 Murcia, Spain.

Department of Fundamental Microbiology, Faculty of Biology and Medicine, University of Lausanne, Quartier UNIL/Sorge, Lausanne, CH1015, Switzerland.

出版信息

Sci Rep. 2017 Feb 24;7:43240. doi: 10.1038/srep43240.

Abstract

CdnL is an essential RNA polymerase (RNAP)-binding activator of rRNA transcription in mycobacteria and myxobacteria but reportedly not in Bacillus. Whether its function and mode of action are conserved in other bacteria thus remains unclear. Because virtually all alphaproteobacteria have a CdnL homolog and none of these have been characterized, we studied the homolog (CdnL) of the model alphaproteobacterium Caulobacter crescentus. We show that CdnL is not essential for viability but that its absence or depletion causes slow growth and cell filamentation. CdnL is degraded in vivo in a manner dependent on its C-terminus, yet excess CdnL resulting from its stabilization did not adversely affect growth. We find that CdnL interacts with itself and with the RNAP β subunit, and localizes to at least one rRNA promoter in vivo, whose activity diminishes upon depletion of CdnL. Interestingly, cells expressing CdnL mutants unable to interact with the RNAP were cold-sensitive, suggesting that CdnL interaction with RNAP is especially required at lower than standard growth temperatures in C. crescentus. Our study indicates that despite limited sequence similarities and regulatory differences compared to its myco/myxobacterial homologs, CdnL may share similar biological functions, since it affects rRNA synthesis, probably by stabilizing open promoter-RNAP complexes.

摘要

CdnL 是分枝杆菌和粘球菌中 rRNA 转录的必需 RNA 聚合酶 (RNAP) 结合激活因子,但据报道在芽孢杆菌中并非如此。因此,其在其他细菌中的功能和作用模式是否保守尚不清楚。由于几乎所有的α-变形菌都有 CdnL 同源物,而这些同源物尚未被描述,我们研究了模式α-变形菌新月柄杆菌的同源物 (CdnL)。我们表明 CdnL 不是生存所必需的,但它的缺失或耗尽会导致生长缓慢和细胞丝状化。CdnL 在体内以依赖于其 C 末端的方式降解,但由于其稳定性而导致的过量 CdnL 不会对生长产生不利影响。我们发现 CdnL 与自身和 RNAPβ亚基相互作用,并在体内定位于至少一个 rRNA 启动子,其活性在 CdnL 耗尽时降低。有趣的是,表达不能与 RNAP 相互作用的 CdnL 突变体的细胞对冷敏感,这表明 CdnL 与 RNAP 的相互作用在新月柄杆菌的低于标准生长温度下尤其需要。我们的研究表明,尽管与分枝杆菌/粘球菌同源物相比,CdnL 的序列相似性和调控差异有限,但 CdnL 可能具有相似的生物学功能,因为它影响 rRNA 合成,可能通过稳定开放启动子-RNAP 复合物来实现。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/49a3/5324124/0399ad60ccc5/srep43240-f1.jpg

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