Gluconeogenic carbon flow of tricarboxylic acid cycle intermediates is critical for Mycobacterium tuberculosis to establish and maintain infection

Metabolic adaptation to the host niche is a defining feature of the pathogenicity ofMycobacterium tuberculosis (Mtb) . In vitro,Mtb is able to grow on a variety of carbon sources, but mounting evidence has implicated fatty acids as the major source of carbon and energy forMtb during infection. When bacterial metabolism is primarily fueled by fatty acids, biosynthesis of sugars from intermediates of the tricarboxylic acid cycle is essential for growth. The role of gluconeogenesis in the pathogenesis ofMtb however remains unaddressed. Phosphoenolpyruvate carboxykinase (PEPCK) catalyzes the first committed step of gluconeogenesis. We applied genetic analyses and13 C carbon tracing to confirm that PEPCK is essential for growth ofMtb on fatty acids and catalyzes carbon flow from tricarboxylic acid cycle–derived metabolites to gluconeogenic intermediates. We further show that PEPCK is required for growth ofMtb in isolated bone marrow–derived murine macrophages and in mice. Importantly,Mtb lacking PEPCK not only failed to replicate in mouse lungs but also failed to survive, and PEPCK depletion during the chronic phase of infection resulted in mycobacterial clearance.Mtb thus relies on gluconeogenesis throughout the infection. PEPCK depletion also attenuatedMtb in IFNγ-deficient mice, suggesting that this enzyme represents an attractive target for chemotherapy.