Glucose and Glycolysis Are Required for the Successful Infection of Macrophages and Mice by Salmonella enterica Serovar Typhimurium

Abstract

Salmonella is a widespread zoonotic enteropathogen that causes gastroenteritis and fatal typhoidal disease in mammals. During systemic infection of mice, Salmonella enterica serovar Typhimurium resides and replicates in macrophages within the "Salmonella-containing vacuole" (SCV). It is surprising that the substrates and metabolic pathways necessary for growth of S. Typhimurium within the SCV of macrophages have not been identified yet. To determine whether S. Typhimurium utilized sugars within the SCV, we constructed a series of S. Typhimurium mutants that lacked genes involved in sugar transport and catabolism and tested them for replication in mice and macrophages. These mutants included a mutant with a mutation in the pfkAB-encoded phosphofructokinase, which catalyzes a key committing step in glycolysis. We discovered that a pfkAB mutant is severely attenuated for replication and survival within RAW 264.7 macrophages. We also show that disruption of the phosphoenolpyruvate:carbohydrate phosphotransferase system by deletion of the ptsHI and crr genes reduces S. Typhimurium replication within RAW 264.7 macrophages. We discovered that mutants unable to catabolize glucose due to deletion of ptsHI, crr, and glk or deletion of ptsG, manXYZ, and glk showed reduced replication within RAW 264.7 macrophages. This study proves that S. Typhimurium requires glycolysis for infection of mice and macrophages and that transport of glucose is required for replication within macrophages.

Glycolysis and pyrimidine biosynthesis are required for replication of adherent–invasive Escherichia coli in macrophages

“…We have analysed the ability of a number of metabolic mutants in AIEC to replicate in macrophages and we have shown that glycolysis has a significant role during growth in the acidified phagosomal niche. This is similar to what has recently been reported for S. enterica serovar Typhimurium where the same mutation (DpfkAB) prevented intracellular replication of S. Typhimurium in murine RAW264.7 macrophages (Bowden et al, 2009). The DpfkAB mutant in Salmonella was also attenuated in virulence and this mutant showed significantly less colonization of BALB/c mice after either intravenous or intraperitoneal injection (Bowden et al, 2009;Paterson et al, 2009).…”

“…This is similar to what has recently been reported for S. enterica serovar Typhimurium where the same mutation (DpfkAB) prevented intracellular replication of S. Typhimurium in murine RAW264.7 macrophages (Bowden et al, 2009). The DpfkAB mutant in Salmonella was also attenuated in virulence and this mutant showed significantly less colonization of BALB/c mice after either intravenous or intraperitoneal injection (Bowden et al, 2009;Paterson et al, 2009). Glycolysis has also been shown to be required for the replication of Shigella within the cytoplasm of infected epithelial cells (Waligora et al, 2014).…”

The Bacterial Phosphoenolpyruvate:Carbohydrate Phosphotransferase System: Regulation by Protein Phosphorylation and Phosphorylation-Dependent Protein-Protein Interactions

“…First, it has been reported that inactivation of the general PTS components EI and HPr prevents the replication of this bacterium in macrophages (130). Second, deletion of EIIA Ntr was found to indirectly affect the virulence of this pathogen.…”

Bacterial peptide transporters: Messengers of nutrition to virulence

“…This may slow down the growth but does not affect survival and therefore leave little scope for the development of resistance. Metabolism associated genes of pathogens have been consistently linked with virulence of major pathogens, [1][2][3][4][5][6][7][8][9] such as, role of carbon metabolism related pathways like glycolysis 10 and TCA cycle 11 in virulence of Salmonella Typhimurium and requirement of metabolism associated pathways for protection of Mycobacterium tuberculosis from host immune system. 9 Salmonella is even known to modulate host to exploit the limited supply of nutrients.…”