The anaplerotic node is essential for the intracellular survival of Mycobacterium tuberculosis

A - Papers appearing in refereed journals

Basu, P., Sandhu, N., Bhatt, A., Singh, A., Balhana, R., Gobe, I., Crowhurst, N. A., Mendum, T. A., Gao, L., Ward, J. L., Beale, M. H., McFadden J. and Beste, D. J. V. 2018. The anaplerotic node is essential for the intracellular survival of Mycobacterium tuberculosis. Journal of Biological Chemistry. 293 (15), pp. 5695-5704. https://doi.org/10.1074/jbc.RA118.001839

AuthorsBasu, P., Sandhu, N., Bhatt, A., Singh, A., Balhana, R., Gobe, I., Crowhurst, N. A., Mendum, T. A., Gao, L., Ward, J. L., Beale, M. H., McFadden J. and Beste, D. J. V.
Abstract

Enzymes at the phosphoenolpyruvate (PEP)–pyruvate–oxaloacetate or anaplerotic (ANA) node control the metabolic flux to glycolysis, gluconeogenesis, and anaplerosis. Here we used genetic, biochemical, and 13C isotopomer analysis to characterize the role of the enzymes at the ANA node in intracellular survival of the world's most successful bacterial pathogen, Mycobacterium tuberculosis (Mtb). We show that each of the four ANA enzymes, pyruvate carboxylase (PCA), PEP carboxykinase (PCK), malic enzyme (MEZ), and pyruvate phosphate dikinase (PPDK), performs a unique and essential metabolic function during the intracellular survival of Mtb. We show that in addition to PCK, intracellular Mtb requires PPDK as an alternative gateway into gluconeogenesis. Propionate and cholesterol detoxification was also identified as an essential function of PPDK revealing an unexpected role for the ANA node in the metabolism of these physiologically important intracellular substrates and highlighting this enzyme as a tuberculosis (TB)-specific drug target. We show that anaplerotic fixation of CO2 through the ANA node is essential for intracellular survival of Mtb and that Mtb possesses three enzymes (PCA, PCK, and MEZ) capable of fulfilling this function. In addition to providing a back-up role in anaplerosis we show that MEZ also has a role in lipid biosynthesis. MEZ knockout strains have an altered cell wall and were deficient in the initial entry into macrophages. This work reveals that the ANA node is a focal point for controlling the intracellular replication of Mtb, which goes beyond canonical gluconeogenesis and represents a promising target for designing novel anti-TB drugs.

KeywordsTuberculosis; Microbial Pathogenesis; Microbiology; Host-pathogen interaction; Mycobacterium tuberculosis; Glucogenesis; Enzyme; Microbial metabolism
Year of Publication2018
JournalJournal of Biological Chemistry
Journal citation293 (15), pp. 5695-5704
Digital Object Identifier (DOI)https://doi.org/10.1074/jbc.RA118.001839
PubMed ID29475946
Web address (URL)http://www.jbc.org/content/293/15/5695
Open accessPublished as ‘gold’ (paid) open access
Publisher's version
Output statusPublished
Publication dates
Online23 Feb 2018
Publication process dates
Accepted23 Feb 2018
PublisherAmerican Society for Biochemistry and Molecular Biology
Copyright licenseCC BY
ISSN0021-9258

Permalink - https://repository.rothamsted.ac.uk/item/84860/the-anaplerotic-node-is-essential-for-the-intracellular-survival-of-mycobacterium-tuberculosis

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