Huang Lige, Hu Xinling, Tao Jun, Mi Kaixia
Wei Sheng Wu Xue Bao. 2014 Nov 4;54(11):1279-88.
Reactive oxygen species are natural products of metabolism in aerobic organisms, which lead to oxidative damage, such as DNA mutation, protein inactivation and drug resistance. MSMEG_3312 was predicted as a hemerythrin-like protein, which can carry oxygen and reversibly bind to oxygen, thus it might play important roles in the process of oxygen metabolism. In this study, we explored the role of MSMEG_3312 in drug resistance.
On the basis of bioinformatics, we identified the conserved sequence of HHE domain in MSMEG_3312 and it was predicted to have typical α-helix at secondary structure. To explore potential functions of MSMEG_3312, we constructed the msmeg_3312 knockout strain and compare the susceptibility to various drugs to its parent strain, mc2155. In addition, we also measured the promoter response when treatment of erythromycin.
Genetic results showed that MSMEG_3312 is not necessary for M. smegmatis growth at 7H9 rich medium. The msmeg_3312 knockout strain showed increased erythromycin resistance. Moreover, the drug resistance is only limited to erythromycin which its mechanism of action is by binding to the 50S subunit of the bacteria ribosomal complex and then inhibit protein synthesis. However, there were no different MICs of other antibiotics, targets for protein synthesis inhibition, but not 50S subunit, such as tetracyclines, aminoglycosides and chloramphenicol. Moreover, we also showed that the promoter of msmeg_3312 responses to erythromycin.
Hemerythin-like protein MSMEG_3312 is involved in erythromycin resistance.
活性氧是需氧生物新陈代谢的天然产物,会导致氧化损伤,如DNA突变、蛋白质失活和耐药性。MSMEG_3312被预测为一种类蚯蚓血红蛋白蛋白,它可以携带氧气并与氧气可逆结合,因此可能在氧代谢过程中发挥重要作用。在本研究中,我们探讨了MSMEG_3312在耐药性中的作用。
基于生物信息学,我们鉴定了MSMEG_3312中HHE结构域的保守序列,并预测其二级结构具有典型的α螺旋。为了探索MSMEG_3312的潜在功能,我们构建了msmeg_3312基因敲除菌株,并比较其与亲本菌株mc2155对各种药物的敏感性。此外,我们还检测了用红霉素处理时的启动子反应。
遗传学结果表明,MSMEG_3312对耻垢分枝杆菌在7H9丰富培养基中的生长不是必需的。msmeg_3312基因敲除菌株对红霉素的耐药性增加。此外,耐药性仅局限于红霉素,其作用机制是通过与细菌核糖体复合物的50S亚基结合,进而抑制蛋白质合成。然而,对于其他抗生素,如四环素、氨基糖苷类和氯霉素,它们的作用靶点是抑制蛋白质合成,但不是50S亚基,其最小抑菌浓度没有差异。此外,我们还表明msmeg_3312的启动子对红霉素有反应。
类蚯蚓血红蛋白蛋白MSMEG_3312参与红霉素耐药性。