School of Life Sciences, Anhui Agricultural University, Hefei, Anhui 230036, China; Institute of Microbe and Host Health, Linyi University, Linyi, Shandong 276005, China.
School of Life Sciences, Anhui Agricultural University, Hefei, Anhui 230036, China.
Poult Sci. 2020 Dec;99(12):6390-6401. doi: 10.1016/j.psj.2020.09.048. Epub 2020 Sep 30.
Avian pathogenic Escherichia coli (APEC) causes a variety of bacterial infectious diseases known as avian colibacillosis leading to significant economic losses in the poultry industry worldwide and restricting the development of the poultry industry. The development of efflux pumps is one important bacterial antibiotic resistance mechanism. Efflux pumps are capable of extruding a wide range of antibiotics out of the cytoplasm of some bacterial species, including β-lactams, polymyxins, tetracyclines, fluoroquinolones, aminoglycosides, novobiocin, nalidixic acid, and fosfomycin. In the present study, we constructed the mcbR mutant and the mcbR-overexpressing strain of E. coli strain APECX40 and performed antimicrobial susceptibility testing, antibacterial activity assays, real-time reverse transcription PCR, and electrophoretic mobility shift assays (EMSA) to investigate the molecular regulatory mechanism of McbR on the genes encoding efflux pumps. Our results showed that McbR positively regulates cell susceptibility to 12 antibiotics, including clindamycin, lincomycin, cefotaxime, cefalexin, doxycycline, tetracycline, gentamicin, kanamycin, norfloxacin, ofloxacin, erythromycin, and rifampicin by activating the transcription of acrAB, acrD, emrD, and mdtD (P < 0.01). Additionally, EMSA indicated that McbR specifically binds to the promoter regions of acrAB, acrD, acrR, emrD, and mdtD. This study suggests that, in APECX40, McbR plays an important role in the regulation of bacterial susceptibility by directly activating the transcription of efflux pumps genes.
禽致病性大肠杆菌(APEC)引起多种细菌性传染病,称为禽大肠杆菌病,导致全球家禽业遭受重大经济损失,并限制了家禽业的发展。外排泵的发展是细菌抗生素耐药性的重要机制之一。外排泵能够将多种抗生素从某些细菌的细胞质中排出,包括β-内酰胺类、多粘菌素类、四环素类、氟喹诺酮类、氨基糖苷类、新生霉素、萘啶酸和磷霉素。在本研究中,我们构建了 mcbR 突变体和 mcbR 过表达的大肠杆菌 APECX40 菌株,并进行了药敏试验、抗菌活性测定、实时逆转录 PCR 和电泳迁移率变动分析(EMSA),以研究 McbR 对编码外排泵基因的分子调控机制。结果表明,McbR 通过激活 acrAB、acrD、emrD 和 mdtD 的转录,正向调节细胞对 12 种抗生素的敏感性,包括克林霉素、林可霉素、头孢噻肟、头孢氨苄、强力霉素、四环素、庆大霉素、卡那霉素、诺氟沙星、氧氟沙星、红霉素和利福平(P<0.01)。此外,EMSA 表明 McbR 特异性结合 acrAB、acrD、acrR、emrD 和 mdtD 的启动子区域。本研究表明,在 APECX40 中,McbR 通过直接激活外排泵基因的转录,在调节细菌敏感性方面发挥重要作用。