Transcriptional analysis of long-term adaptation of Yersinia enterocolitica to low-temperature growth
Transcriptional analysis of long-term adaptation of Yersinia enterocolitica to low-temperature growth
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DOI：
10.1128/jb.188.8.2945-2958.2006
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发表时间：
2006-04-01
影响因子：
3.2
通讯作者：
Fuchs, TM
中科院分区：
文献类型：
--
作者：
Bresolin, G;Neuhaus, K;Fuchs, TM
为了分析小肠结肠炎耶尔森氏菌细胞对低温下延长生长的转录应答，在最佳（30 ° C）和次最佳（10 ° C）温度下平行培养luxCDABE转座子突变体的集合，并在生长期间筛选增强的启动子活性，直到进入稳定期。在5，700 Y.在小肠结肠炎菌突变体中，鉴定了42个转录单位，与30 ℃相比，10 ℃下启动子活性强烈增强或降低，并测量了它们转录水平随时间的变化。与10个启动子区域融合的绿色荧光蛋白证实了该数据。Y.温度反应基因的诱导时间顺序。从冷休克基因cspA和cspB的表达以及谷氨酸-天冬氨酸同向转运体的转录升高开始，推断小肠结肠炎。随后，冷适应细胞大幅上调编码环境传感器和调节因子的基因，例如UhpABC、ArcA和甲基接受趋化蛋白I（MCPI）。其中最突出的冷响应元件，在早期和中期指数期的增长过程中转录诱导的杀虫毒素基因tcaA和tcaB，以及参与鞭毛合成和趋化性的基因。晚期指数生长期到早期稳定生长期的表达模式由参与生物降解代谢的因素主导，即组氨酸氨裂解酶、负责糖原摄取和利用的三种酶、尿素酶复合物和枯草杆菌蛋白酶样蛋白酶。双敲除突变体和互补研究表明MCPI和UhpC对推定的溶血素转运蛋白表达的抑制作用。这些数据部分地描绘了Y.在环境温度下的小肠结肠炎，提供证据表明，一个迄今未知的昆虫阶段是这种人类病原体的生命周期的一部分。
To analyze the transcriptional response of Yersinia enterocolitica cells to prolonged growth at low temperature, a collection of luxCDABE transposon mutants was cultivated in parallel at optimal (30 degrees C) and suboptimal (10 degrees C) temperatures and screened for enhanced promoter activities during growth until entering stationary phase. Among 5,700 Y. enterocolitica mutants, 42 transcriptional units were identified with strongly enhanced or reduced promoter activity at 10 degrees C compared to 30 degrees C, and changes in their transcriptional levels over time were measured. Green fluorescent protein fusions to 10 promoter regions confirmed the data. The temporal order of induction of the temperature-responsive genes of Y. enterocolitica was deduced, starting with the expression of cold shock genes cspA and cspB and the elevated transcription of a glutamate-aspartate symporter. Subsequently, cold-adapted cells drastically up-regulated genes encoding environmental sensors and regulators, such as UhpABC, ArcA, and m ethyl-accepting chemotaxis protein I (MCPI). Among the most prominent cold-responsive elements that were transcriptionally induced during growth in early and middle exponential phase are the insecticidal toxin genes tcaA and tcaB, as well as genes involved in flagellar synthesis and chemotaxis. The expression pattern of the late-exponential- to early-stationary-growth phase is dominated by factors involved in biodegradative metabolism, namely, a histidine ammonia lyase, three enzymes responsible for uptake and utilization of glycogen, the urease complex, and a subtilisin-like protease. Double-knockout mutants and complementation studies demonstrate inhibitory effects of MCPI and UhpC on the expression of a putative hemolysin transporter. The data partially delineate the spectrum of gene expression of Y. enterocolitica at environmental temperatures, providing evidence that an as-yet-unknown insect phase is part of the life cycle of this human pathogen.