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Integrated Transcriptomic and Proteomic Analysis of the Physiological Response of Escherichia coli O157:H7 Sakai to Steady-state Conditions of Cold and Water Activity Stress

机译:大肠杆菌O157:H7 Sakai对冷水和水分活动应激稳态条件生理响应的转录组和蛋白质组学综合分析

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摘要

An integrated transcriptomic and proteomic analysis was undertaken to determine the physiological response of Escherichia coli O157:H7 Sakai to steady-state conditions relevant to low temperature and water activity conditions experienced during meat carcass chilling in cold air. The response of E. coli during exponential growth at 25 °C aw 0.985, 14 °C aw 0.985, 25 °C aw 0.967, and 14 °C aw 0.967 was compared with that of a reference culture (35 °C aw 0.993). Gene and protein expression profiles of E. coli were more strongly affected by low water activity (aw 0.967) than by low temperature (14 °C). Predefined group enrichment analysis revealed that a universal response of E. coli to all test conditions included activation of the master stress response regulator RpoS and the Rcs phosphorelay system involved in the biosynthesis of the exopolysaccharide colanic acid, as well as down-regulation of elements involved in chemotaxis and motility. However, colanic acid-deficient mutants were shown to achieve comparable growth rates to their wild-type parents under all conditions, indicating that colanic acid is not required for growth. In contrast to the transcriptomic data, the proteomic data revealed that several processes involved in protein synthesis were down-regulated in overall expression at 14 °C aw 0.985, 25 °C aw 0.967, and 14 °C aw 0.967. This result suggests that during growth under these conditions, E. coli, although able to transcribe the required mRNA, may lack the cellular resources required for translation. Elucidating the global adaptive response of E. coli O157:H7 during exposure to chilling and water activity stress has provided a baseline of knowledge of the physiology of this pathogen.
机译:进行了综合的转录组和蛋白质组学分析,以确定大肠杆菌O157:H7 Sakai对与在冷空气中冷冻肉meat体时经历的低温和水分活度条件相关的稳态条件的生理反应。将大肠杆菌在25°C的Aw 0.985、14°C的Aw 0.985、25°C的Aw 0.967和14°C的Aw 0.967指数生长期间的响应与参考培养物(35°C的Aw 0.993)进行了比较。低水分活度(aw 0.967)比低温(14°C)对大肠杆菌的基因和蛋白质表达谱的影响更大。预先定义的组富集分析表明,大肠杆菌对所有测试条件的普遍反应包括激活激活的主应激反应调节剂RpoS和参与胞外多糖可乐酸生物合成的Rcs磷酸化系统,以及所涉及元素的下调在趋化性和运动性。但是,在所有条件下,均可证明缺少缺角菌酸的突变体可达到与其野生型亲本相当的生长速率,这表明不需要新增缺菌酸。与转录组数据相反,蛋白质组学数据揭示了蛋白质合成中涉及的几个过程在14°C aw 0.985、25°C aw 0.967和14°C aw 0.967时的总表达下调。该结果表明,在这些条件下的生长过程中,大肠杆菌虽然能够转录所需的mRNA,但可能缺少翻译所需的细胞资源。阐明大肠杆菌O157:H7在暴露于寒冷和水分活动压力期间的总体适应性反应为该病原体的生理知识提供了基线。

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