Pöntinen Anna, Markkula Annukka, Lindström Miia, Korkeala Hannu
Department of Food Hygiene and Environmental Health, Faculty of Veterinary Medicine, University of Helsinki, Helsinki, Finland
Department of Food Hygiene and Environmental Health, Faculty of Veterinary Medicine, University of Helsinki, Helsinki, Finland.
Appl Environ Microbiol. 2015 Jun 15;81(12):3994-4004. doi: 10.1128/AEM.00626-15. Epub 2015 Apr 3.
Two-component systems (TCSs) aid bacteria in adapting to a wide variety of stress conditions. While the role of TCS response regulators in the cold tolerance of the psychrotrophic foodborne pathogen Listeria monocytogenes has been demonstrated previously, no comprehensive studies showing the role of TCS histidine kinases of L. monocytogenes at low temperature have been performed. We compared the expression levels of each histidine kinase-encoding gene of L. monocytogenes EGD-e in logarithmic growth phase at 3°C and 37°C, as well as the expression levels 30 min, 3 h, and 7 h after cold shock at 5°C and preceding cold shock (at 37°C). We constructed a deletion mutation in each TCS histidine kinase gene, monitored the growth of the EGD-e wild-type and mutant strains at 3°C and 37°C, and measured the minimum growth temperature of each strain. Two genes, yycG and lisK, proved significant in regard to induced relative expression levels under cold conditions and cold-sensitive mutant phenotypes. Moreover, the ΔresE mutant showed a lower growth rate than that of wild-type EGD-e at 3°C. Eleven other genes showed upregulated gene expression but revealed no cold-sensitive phenotypes. The results show that the histidine kinases encoded by yycG and lisK are important for the growth and adaptation of L. monocytogenes EGD-e at low temperature.
双组分系统(TCSs)帮助细菌适应各种各样的应激条件。虽然之前已经证明了TCS反应调节因子在嗜冷食源性病原体单核细胞增生李斯特菌的耐寒性中的作用,但尚未进行全面研究来表明单核细胞增生李斯特菌的TCS组氨酸激酶在低温下的作用。我们比较了单核细胞增生李斯特菌EGD-e在3°C和37°C对数生长期时每个组氨酸激酶编码基因的表达水平,以及在5°C冷休克30分钟、3小时和7小时后以及冷休克前(37°C)的表达水平。我们在每个TCS组氨酸激酶基因中构建了缺失突变,监测了EGD-e野生型和突变株在3°C和37°C下的生长情况,并测量了每个菌株的最低生长温度。结果证明,yycG和lisK这两个基因在低温条件下的诱导相对表达水平和冷敏感突变体表型方面具有重要意义。此外,ΔresE突变体在3°C时的生长速率低于野生型EGD-e。其他11个基因显示基因表达上调,但未表现出冷敏感表型。结果表明,yycG和lisK编码的组氨酸激酶对单核细胞增生李斯特菌EGD-e在低温下的生长和适应至关重要。