Inactivation of fructose-1,6-bisphosphate aldolase prevents optimal co-catabolism of glycolytic and gluconeogenic carbon substrates in Mycobacterium tuberculosis.
Metabolic pathways used by Mycobacterium tuberculosis (Mtb) to establish and maintain infections are important for our understanding of pathogenesis and the development of new chemotherapies. To investigate the role of fructose-1,6-bisphosphate aldolase (FBA), we engineered an Mtb strain in which FBA levels were regulated by anhydrotetracycline. Depletion of FBA resulted in clearance of Mtb in both the acute and chronic phases of infection in vivo, and loss of viability in vitro when cultured on single carbon sources. Consistent with prior reports of Mtb's ability to co-catabolize multiple carbon sources, this in vitro essentiality could be overcome when cultured on mixtures of glycolytic and gluconeogenic carbon sources, enabling generation of an fba knockout (Δfba). In vitro studies of Δfba however revealed that lack of FBA could only be compensated for by a specific balance of glucose and butyrate in which growth and metabolism of butyrate were determined by Mtb's ability to co-catabolize glucose. These data thus not only evaluate FBA as a potential drug target in both replicating and persistent Mtb, but also expand our understanding of the multiplicity of in vitro conditions that define the essentiality of Mtb's FBA in vivo.
Department of Microbiology and Immunology, Weill Cornell Medical College, New York, New York, United States of America.
Susan
S
Puckett
Department of Microbiology and Immunology, Weill Cornell Medical College, New York, New York, United States of America.
Carolina
C
Trujillo
Department of Medicine, Weill Cornell Medical College, New York, New York, United States of America.
Hyungjin
H
Eoh
Department of Microbiology and Immunology, Weill Cornell Medical College, New York, New York, United States of America.
Joeli
J
Marrero
Mycobacteria Research Laboratories, Department of Microbiology, Immunology and Pathology, Colorado State University, Fort Collins, Colorado, United States of America.
John
J
Spencer
Mycobacteria Research Laboratories, Department of Microbiology, Immunology and Pathology, Colorado State University, Fort Collins, Colorado, United States of America.
Mary
M
Jackson
Department of Microbiology and Immunology, Weill Cornell Medical College, New York, New York, United States of America.
Dirk
D
Schnappinger
Department of Medicine, Weill Cornell Medical College, New York, New York, United States of America.
Kyu
K
Rhee
Department of Microbiology and Immunology, Weill Cornell Medical College, New York, New York, United States of America.
Sabine
S
Ehrt
D000818:Animals; D002087:Butyrates / Q000378:metabolism; D050260:Carbohydrate Metabolism / Q000235:genetics; D005260:Female; D005634:Fructose-Bisphosphate Aldolase / Q000235:genetics*; D017353:Gene Deletion; D005943:Gluconeogenesis / Q000235:genetics*; D006019:Glycolysis / Q000235:genetics*; D008660:Metabolism / Q000235:genetics; D055442:Metabolome / Q000235:genetics; D051379:Mice; D008810:Mice, Inbred C57BL; D009169:Mycobacterium tuberculosis / Q000235:genetics* / Q000378:metabolism*; D030781:Organisms, Genetically Modified
D002087:Butyrates; D005634:Fructose-Bisphosphate Aldolase