Hu Y M, Butcher P D, Sole K, Mitchison D A, Coates A R
Department of Medical Microbiology, St. George's Hospital Medical School, London, United Kingdom.
FEMS Microbiol Lett. 1998 Jan 1;158(1):139-45. doi: 10.1111/j.1574-6968.1998.tb12813.x.
Oxygen-limiting conditions are critical to the survival of the bacteria in tuberculosis. Mycobacterium tuberculosis can survive anaerobiosis in vitro for long periods of time only after a gradual transition to a microaerophilic stationary phase. The underlying mechanism behind stationary phase adaption needs to be elucidated. The protein profiles of Mycobacterium tuberculosis during long-term stationary phase growth and under strict anaerobic incubation were monitored by [35S]methionine labelling, SDS-PAGE and fluorography. These experiments have established that protein synthesis gradually decreased over 50 days in the long-term stationary phase cultures which were considered to be microaerophilic. There was an 80% linear decrease in the level of total protein synthesis during the first 40 days of microaerophilic growth and then the rate of protein synthesis faded quickly. For the first time we have shown that total protein synthesis shutdown occurred when bacilli were incubated under further anaerobic conditions. Viability, estimated by cfu counts, remained constant during stationary phase growth and under anaerobic incubation. Furthermore, when oxygen was introduced into the anaerobic culture, protein synthesis restarted. Also heat shock at 45 degrees C, 48 degrees C and 50 degrees C rapidly induced protein synthesis in stationary and anaerobic cultures. These data indicate that dormant bacteria shut down protein synthesis but remain responsive to specific stimuli which restore protein synthesis. In addition the dormant bacilli induced by anaerobiosis developed more heat resistance than nondormant organisms.
氧气限制条件对结核分枝杆菌的存活至关重要。结核分枝杆菌只有在逐渐过渡到微需氧稳定期后,才能在体外长时间厌氧存活。稳定期适应背后的潜在机制有待阐明。通过[35S]甲硫氨酸标记、SDS-PAGE和荧光自显影监测结核分枝杆菌在长期稳定期生长和严格厌氧培养条件下的蛋白质谱。这些实验表明,在被认为是微需氧的长期稳定期培养物中,蛋白质合成在50天内逐渐减少。在微需氧生长的前40天,总蛋白质合成水平呈80%的线性下降,然后蛋白质合成速率迅速下降。我们首次表明,当杆菌在进一步厌氧条件下培养时,总蛋白质合成停止。通过菌落形成单位计数估计的活力在稳定期生长和厌氧培养期间保持恒定。此外,当向厌氧培养物中引入氧气时,蛋白质合成重新开始。同样,在45℃、48℃和50℃下的热休克迅速诱导稳定期和厌氧培养物中的蛋白质合成。这些数据表明,休眠细菌停止蛋白质合成,但仍对恢复蛋白质合成的特定刺激有反应。此外,由厌氧诱导的休眠杆菌比非休眠生物体具有更强的耐热性。