Insight into the role of a novel c-di-GMP effector protein in Rhodococcus ruber.
Insight into the role of a novel c-di-GMP effector protein in Rhodococcus ruber.
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DOI：
10.1016/j.bbrc.2022.03.131
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发表时间：
2022-03
影响因子：
3.1
通讯作者：
Yuan Yuan-Yuan;Fan Zhang;Lei Ai;Yuzhu Huang;R. Peng
中科院分区：
文献类型：
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作者：
Yuan Yuan-Yuan;Fan Zhang;Lei Ai;Yuzhu Huang;R. Peng
c -二gmp是细菌中普遍存在的第二信使，调控生物膜形成、细胞迁移、毒力、细胞粘附、细胞周期等细胞功能。这些功能与c-二gmp效应蛋白和/或核糖体开关数量的增加有关。本研究采用亲和下拉和LC/MS/MS相结合的方法筛选了一种新的c-di-GMP结合蛋白CEP1 （c-di-GMP效应蛋白1）。表面等离子体共振证实了CEP1与c-di-GMP的结合，解离常数为127±1.03 μM。实时荧光定量PCR结果显示，经甲苯和苯酚胁迫后，红球菌SD3中cep1基因的mRNA表达量分别增加了63.29倍和71.18倍。利用穿梭质粒pNV18构建cep1基因增强菌株，在不同有机溶剂存在下，cep1基因增强菌株的生长均优于野生型菌株。本研究揭示了c-di-GMP与红球菌SD3耐有机溶剂性相关的机制。
C-di-GMP is a ubiquitous second messenger in bacterium, which regulates cellular functions such as the formation of biofilm membrane, cell mobility, virulence, cell adhesion, cell cycle et al. These functions are associated with an increasing number of c-di-GMP effector proteins and/or riboswitchs. In the study, CEP1 (c-di-GMP effector protein 1), a novel c-di-GMP binding protein, was screened with a combination of affinity pull-down and LC/MS/MS methods. The binding of CEP1 and c-di-GMP was demonstrated by surface plasmon resonance, with the dissociation constants of 127 ± 1.03 μM. Quantitative real time PCR assay showed the mRNA levels of cep1 gene in Rhodococcusruber SD3 increased to 63.29 times and 71.18 times after toluene and phenol stress, respectively. Furthermore, cep1 gene enhanced strain was constructed using shuttle plasmid pNV18, which showed improved growth compared to the wild-type strain in the presence of different organic solvents. The study provided an insight into a mechanism, by which c-di-GMP was connected with organic solvent tolerance of Rhodococcusruber SD3.