DI Yuchang, Bai Jiacheng, Chi Mingzhe, Fan Weixing, Zhang Xuelian
State Key Laboratory of Genetic Engineering, School of Life Sciences, Fudan University, Shanghai 200433, China.
Laboratory of Zoonosis, China Animal Health and Epidemiology Center, Qingdao 266032, Shangdong, China.
Sheng Wu Gong Cheng Xue Bao. 2022 Mar 25;38(3):1050-1060. doi: 10.13345/j.cjb.210207.
As the only translational factor that plays a critical role in two translational processes (elongation and ribosome regeneration), GTPase elongation factor G (EF-G) is a potential target for antimicrobial agents. Both and have two EF-G homologous coding genes, EFG1 () and EFG2 (), () and (), respectively. EFG1 () and () were identified as essential genes for bacterial growth by gene mutation library and bioinformatic analysis. To investigate the biological function and characteristics of EF-G in mycobacterium, two induced EF-G knockdown strains (Msm-ΔEFG1(KD) and Msm-ΔEFG2(KD)) from were constructed by clustered regularly interspaced short palindromic repeats interference (CRISPRi) technique. EF-G2 knockdown had no effect on bacterial growth, while EF-G1 knockdown significantly retarded the growth of mycobacterium, weakened the film-forming ability, changed the colony morphology, and increased the length of mycobacterium. It was speculated that EF-G might be involved in the division of bacteria. Minimal inhibitory concentration assay showed that inhibition of EF-G1 expression enhanced the sensitivity of mycobacterium to rifampicin, isoniazid, erythromycin, fucidic acid, capreomycin and other antibacterial agents, suggesting that EF-G1 might be a potential target for screening anti-tuberculosis drugs in the future.
作为唯一在两个翻译过程(延伸和核糖体再生)中起关键作用的翻译因子,GTPase延伸因子G(EF-G)是抗菌药物的潜在靶点。 和 分别有两个EF-G同源编码基因,即EFG1( )和EFG2( ), ( )和 ( )。通过基因突变文库和生物信息学分析,EFG1( )和 ( )被鉴定为细菌生长的必需基因。为了研究分枝杆菌中EF-G的生物学功能和特性,利用成簇规律间隔短回文重复序列干扰(CRISPRi)技术构建了来自 的两个诱导型EF-G敲低菌株(Msm-ΔEFG1(KD)和Msm-ΔEFG2(KD))。敲低EF-G2对细菌生长没有影响,而敲低EF-G1则显著抑制分枝杆菌的生长,削弱其成膜能力,改变菌落形态,并增加分枝杆菌的长度。推测EF-G可能参与细菌的分裂。最低抑菌浓度测定表明,抑制EF-G1的表达增强了分枝杆菌对利福平、异烟肼、红霉素、夫西地酸、卷曲霉素等抗菌药物的敏感性,提示EF-G1可能是未来筛选抗结核药物的潜在靶点。