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

Basu, Piyali, Sandhu, Noor, Bhatt, Apoorva, Singh, Albel, Balhana, Ricardo, Gobe, Irene, Crowhurst, Nicola A, Mendum, Thomas, Gao, Liang, Ward, Jane L , Beale, Michael H, McFadden, Johnjoe and Beste, Dany (2018) The anaplerotic node is essential for the intracellular survival of Mycobacterium tuberculosis Journal of Biological Chemistry, 293 (15). pp. 5695-5704.

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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.

Item Type: Article
Divisions : Faculty of Health and Medical Sciences > School of Biosciences and Medicine
Authors :
NameEmailORCID
Basu, Piyali
Sandhu, Noor
Bhatt, Apoorva
Singh, Albel
Balhana, Ricardo
Gobe, Irene
Crowhurst, Nicola A
Mendum, ThomasT.Mendum@surrey.ac.uk
Gao, Liang
Ward, Jane L
Beale, Michael H
McFadden, JohnjoeJ.Mcfadden@surrey.ac.uk
Beste, DanyD.Beste@surrey.ac.uk
Date : 23 February 2018
Funders : MRC; BBSRC
DOI : 10.1074/jbc.RA118.001839
Copyright Disclaimer : © 2018 Basu et al. Published by The American Society for Biochemistry and Molecular Biology, Inc. Final version open access under the terms of the Creative Commons CC-BY license
Depositing User : Melanie Hughes
Date Deposited : 26 Jul 2018 08:18
Last Modified : 11 Dec 2018 11:24
URI: http://epubs.surrey.ac.uk/id/eprint/848791

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