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首页> 外文期刊>PLoS Genetics >Use of a microfluidic platform to uncover basic features of energy and environmental stress responses in individual cells of Bacillus subtilis
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Use of a microfluidic platform to uncover basic features of energy and environmental stress responses in individual cells of Bacillus subtilis

机译：使用微流体平台揭示枯草芽孢杆菌单个细胞中能量和环境胁迫反应的基本特征

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摘要

Bacteria use a variety of stress-sensing systems to sense and respond to diverse stressors and to ensure their survival under adverse conditions. The gram-positive bacterium Bacillus subtilis responds to energy stress (ATP depletion) and to environmental stressors using two distinct stress-sensing pathways that converge on the alternative sigma factor σB to provoke a general stress response. Past efforts to study the σB stress response in bulk culture and on agarose pads were unable to visualize the responses of individual cells under tightly controlled conditions for extended periods of time. Here we use a microfluidics-based strategy to discern the basic features of σB activation in single cells in response to energy and environmental stress, both immediately upon stressor exposure and for tens of generations thereafter. Upon energy stress at various levels of stressor, cells exhibited fast, transient, and amplitude-modulated responses but not frequency modulation as previously reported. Upon environmental stress, which is mediated by the stressosome complex, wild-type cells primarily exhibited a transient and amplitude-modulated response. However, mutant cells producing only one of the four paralogous RsbR stressosome proteins showed striking and previously unseen differences. Whereas RsbRA-only cells mimicked the wild type, RsbRC-only cells displayed a slower but sustained overall response composed of repeated activation events in single cells.
机译：细菌使用各种压力感应系统来感应和响应各种压力源，并确保其在不利条件下的生存。革兰氏阳性枯草芽孢杆菌利用两种截然不同的压力感应途径对能量压力（ATP耗竭）和环境压力源做出反应，这两种途径在可替代的σ因子σB上收敛，从而引起总体压力响应。过去在大量培养和琼脂糖垫上研究σB应激反应的努力无法在严格控制的条件下长时间观察单个细胞的反应。在这里，我们使用基于微流体的策略来识别能量和环境压力下单个细胞中σB激活的基本特征，既可以在应力暴露后立即使用，也可以在此后的数十代中使用。在各种压力源的能量压力下，细胞表现出快速，瞬时和幅度调制的响应，但没有频率调制，如先前报道的那样。在由应激体复合物介导的环境胁迫下，野生型细胞主要表现出瞬时的和幅度调节的响应。但是，仅产生四个旁源RsbR应激体蛋白之一的突变细胞显示出惊人的差异，并且以前是看不见的。仅有RsbRA的细胞模仿野生型，而仅有RsbRC的细胞显示出较慢但持续的总体反应，该反应由单个细胞中的重复激活事件组成。

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