Bowden Steven D, Hopper-Chidlaw Amanda C, Rice Christopher J, Ramachandran Vinoy K, Kelly David J, Thompson Arthur
Institute of Food Research, Norwich Research Park, Colney, Norwich, United Kingdom; Graduate School of Biological Sciences, Nara Institute of Science and Technology, Ikoma, Nara, Japan.
Institute of Food Research, Norwich Research Park, Colney, Norwich, United Kingdom.
PLoS One. 2014 May 5;9(5):e96266. doi: 10.1371/journal.pone.0096266. eCollection 2014.
Salmonella is the causative agent of a spectrum of human and animal diseases ranging from gastroenteritis to typhoid fever. It is a food--and water--borne pathogen and infects via ingestion followed by invasion of intestinal epithelial cells and phagocytic cells. In this study we employed a mutational approach to define the nutrients and metabolic pathways required by Salmonella enterica serovar Typhimurium during infection of a human epithelial cell line (HeLa). We deleted the key glycolytic genes, pfkA and pfkB to show that S. Typhimurium utilizes glycolysis for replication within HeLa cells; however, glycolysis was not absolutely essential for intracellular replication. Using S. Typhimurium strains deleted for genes encoding components of the phosphotransferase system and glucose transport, we show that glucose is a major substrate required for the intracellular replication of S. Typhimurium in HeLa cells. We also deleted genes encoding enzymes involved in the utilization of gluconeogenic substrates and the glyoxylate shunt and show that neither of these pathways were required for intracellular replication of S. Typhimurium within HeLa cells.
沙门氏菌是一系列人类和动物疾病的病原体,这些疾病从肠胃炎到伤寒热不等。它是一种通过食物和水传播的病原体,通过摄入感染,随后侵入肠道上皮细胞和吞噬细胞。在本研究中,我们采用突变方法来确定鼠伤寒沙门氏菌在感染人上皮细胞系(HeLa)期间所需的营养物质和代谢途径。我们删除了关键的糖酵解基因pfkA和pfkB,以表明鼠伤寒沙门氏菌利用糖酵解在HeLa细胞内进行复制;然而,糖酵解对于细胞内复制并非绝对必要。使用缺失编码磷酸转移酶系统和葡萄糖转运组件基因的鼠伤寒沙门氏菌菌株,我们表明葡萄糖是鼠伤寒沙门氏菌在HeLa细胞内复制所需的主要底物。我们还删除了编码参与糖异生底物利用和乙醛酸循环酶的基因,并表明这些途径对于鼠伤寒沙门氏菌在HeLa细胞内的细胞内复制都不是必需的。