title: Fumarase Deficiency Causes Protein and Metabolite Succination and Intoxicates Mycobacterium tuberculosis.
abstract: Enzymes of central carbon metabolism are essential mediators of Mycobacterium tuberculosis (Mtb) physiology and pathogenicity, but are often perceived to lack sufficient species selectivity to be pursued as potential drug targets. Fumarase (Fum) is an enzyme of the canonical tricarboxylic acid cycle and is dispensable in many organisms. Transposon mutagenesis studies in Mtb, however, indicate that Fum is required for optimal growth. Here, we report the generation and characterization of a genetically engineered Mtb strain in which Fum expression is conditionally regulated. This revealed that Fum deficiency is bactericidal in vitro and during both the acute and chronic phases of mouse infection. This essentiality is linked to marked accumulations of fumarate resulting in protein and metabolite succination, a covalent modification of cysteine thiol residues. These results identify Mtb Fum as a potentially species-specific drug target whose inactivation may kill Mtb through a covalently irreversible form of metabolic toxicity.
authors.affiliation: Department of Microbiology and Immunology, Weill Cornell Medical College, New York, NY 10065, USA.
authors.forename: Nadine
authors.initials: N
authors.lastname: Ruecker
authors.affiliation: Department of Medicine, Weill Cornell Medical College, New York, NY 10065, USA.
authors.forename: Robert
authors.initials: R
authors.lastname: Jansen
authors.affiliation: Department of Microbiology and Immunology, Weill Cornell Medical College, New York, NY 10065, USA.
authors.forename: Carolina
authors.initials: C
authors.lastname: Trujillo
authors.affiliation: Department of Microbiology and Immunology, Weill Cornell Medical College, New York, NY 10065, USA.
authors.forename: Susan
authors.initials: S
authors.lastname: Puckett
authors.affiliation: Department of Microbiology and Immunology, Weill Cornell Medical College, New York, NY 10065, USA.
authors.forename: Pradeepa
authors.initials: P
authors.lastname: Jayachandran
authors.affiliation: Department of Pharmacology, Physiology & Neuroscience, School of Medicine, University of South Carolina, Columbia, SC 29209, USA.
authors.forename: Gerardo G
authors.initials: GG
authors.lastname: Piroli
authors.affiliation: Department of Pharmacology, Physiology & Neuroscience, School of Medicine, University of South Carolina, Columbia, SC 29209, USA.
authors.forename: Norma
authors.initials: N
authors.lastname: Frizzell
authors.affiliation: Proteomics Resource Center, Rockefeller University, New York, NY 10065, USA.
authors.forename: Henrik
authors.initials: H
authors.lastname: Molina
authors.affiliation: Department of Medicine, Weill Cornell Medical College, New York, NY 10065, USA.
authors.forename: Kyu Y
authors.initials: KY
authors.lastname: Rhee
authors.affiliation: Department of Microbiology and Immunology, Weill Cornell Medical College, New York, NY 10065, USA. Electronic address: sae2004@med.cornell.edu.
authors.forename: Sabine
authors.initials: S
authors.lastname: Ehrt
mesh_terms: D000818:Animals; D001426:Bacterial Proteins / Q000235:genetics* / Q000378:metabolism; D002851:Chromatography, High Pressure Liquid; D002952:Citric Acid Cycle; D003545:Cysteine / Q000737:chemistry; D005260:Female; D005649:Fumarate Hydratase / Q000172:deficiency / Q000235:genetics* / Q000378:metabolism; D005650:Fumarates / Q000032:analysis / Q000378:metabolism; D051379:Mice; D008810:Mice, Inbred C57BL; D009169:Mycobacterium tuberculosis / Q000201:enzymology / Q000235:genetics* / Q000472:pathogenicity; D018384:Oxidative Stress; D010455:Peptides / Q000032:analysis / Q000737:chemistry; D011499:Protein Processing, Post-Translational; D053719:Tandem Mass Spectrometry
chemical_list: D001426:Bacterial Proteins; D005650:Fumarates; D010455:Peptides; D005649:Fumarate Hydratase; D003545:Cysteine
keywords: central carbon metabolism; drug target; fumarase; succination; tuberculosis