Zinc-dependent substrate-level phosphorylation powers Salmonella growth under nitrosative stress of the innate host response

The metabolic processes that enable the replication of intracellular Salmonella under nitrosative stress conditions engendered in the innate response of macrophages are poorly understood. A screen of Salmonella transposon mutants identified the ABC-type high-affinity zinc uptake system ZnuABC as a critical determinant of the adaptation of Salmonella to the nitrosative stress generated by the enzymatic activity of inducible nitric oxide (NO) synthase of mononuclear phagocytic cells. NO limits the virulence of a znuB mutant in an acute murine model of salmonellosis. The ZnuABC transporter is crucial for the glycolytic function of fructose bisphosphate aldolase, thereby fueling growth of Salmonella during nitrosative stress produced in the innate response of macrophages. Our investigations demonstrate that glycolysis mediates resistance of Salmonella to the antimicrobial activity of NO produced in an acute model of infection. The ATP synthesized by substrate-level phosphorylation at the payoff phase of glycolysis and acetate fermentation powers the replication of Salmonella experiencing high levels of nitrosative stress. In contrast, despite its high potential for ATP synthesis, oxidative phosphorylation is a major target of inhibition by NO and contributes little to the antinitrosative defenses of intracellular Salmonella. Our investigations have uncovered a previously unsuspected conjunction between zinc homeostasis, glucose metabolism and cellular energetics in the adaptation of intracellular Salmonella to the reactive nitrogen species synthesized in the innate host response.
Author summary Microbial pathogens are exposed to multiple antimicrobial defenses during their associations with host cells. Nitric oxide generated in the innate response exerts widespread antimicrobial activity against a variety of pathogenic microorganisms. Nitric oxide has high affinity for metal groups of terminal cytochromes of the respiratory chain, and thus nitrosative stress exerts extreme deleterious actions against the cellular energetics that rely on oxidative phosphorylation. Intracellular Salmonella have resolved this dilemma by satisfying a significant portion of their energetic demands via substrate level phosphorylation in the payoff phase of glycolysis and acetate fermentation. A high affinity zinc uptake system promotes antinitrosative defense of intracellular Salmonella by in great part supporting the enzymatic activity of an essential enzyme in the preparatory phase of glycolysis. Our research provides novel insights into the metabolic and energetic adaptations that allow a bacterial pathogen to thrive in the midst of the innate host response of vertebrate cells.