Deletion of transketolase triggers a stringent metabolic response in promastigotes and loss of virulence in amastigotes of Leishmania mexicana

Kovarova, J., Pountain, A. W. , Wildridge, D., Weidt, S., Bringaud, F., Burchmore, R. J.S. , Achcar, F. and Barrett, M. P. (2018) Deletion of transketolase triggers a stringent metabolic response in promastigotes and loss of virulence in amastigotes of Leishmania mexicana. PLoS Pathogens, 14(3), e1006953. (doi: 10.1371/journal.ppat.1006953) (PMID:29554142) (PMCID:PMC5882173)

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Transketolase (TKT) is part of the non-oxidative branch of the pentose phosphate pathway (PPP). Here we describe the impact of removing this enzyme from the pathogenic protozoan Leishmania mexicana. Whereas the deletion had no obvious effect on cultured promastigote forms of the parasite, the Δtkt cells were not infective to mice. Δtkt promastigotes were more susceptible to oxidative stress and various leishmanicidal drugs than wild-type, and metabolomics analysis revealed profound changes to metabolism in these cells. In addition to changes consistent with those directly related to the role of TKT in the PPP, central carbon metabolism was substantially decreased, the cells consumed significantly less glucose, flux through glycolysis diminished, and production of the main end products of metabolism was decreased. Only minor changes in RNA abundance from genes encoding enzymes in central carbon metabolism, however, were detected although fructose-1,6-bisphosphate aldolase activity was decreased two-fold in the knock-out cell line. We also showed that the dual localisation of TKT between cytosol and glycosomes is determined by the C-terminus of the enzyme and by engineering different variants of the enzyme we could alter its sub-cellular localisation. However, no effect on the overall flux of glucose was noted irrespective of whether the enzyme was found uniquely in either compartment, or in both.

Item Type:Articles
Glasgow Author(s) Enlighten ID:Burchmore, Dr Richard and Weidt, Dr Stefan and Achcar, Dr Fiona and Pountain, Andrew and Kovarova, Ms Julie and Barrett, Professor Michael
Creator Roles:
Kovářová, J.Conceptualization, Formal analysis, Investigation, Validation, Visualization, Writing – original draft, Writing – review and editing
Pountain, A. W.Investigation, Methodology, Validation, Writing – original draft
Weidt, S.Data curation, Formal analysis, Methodology
Burchmore, R. J.S.Conceptualization, Funding acquisition, Project administration, Supervision
Achcar, F.Conceptualization, Data curation, Formal analysis
Barrett, M. P.Conceptualization, Funding acquisition, Project administration, Resources, Supervision, Writing – original draft, Writing – review and editing
Authors: Kovarova, J., Pountain, A. W., Wildridge, D., Weidt, S., Bringaud, F., Burchmore, R. J.S., Achcar, F., and Barrett, M. P.
College/School:College of Medical Veterinary and Life Sciences > School of Infection & Immunity
Journal Name:PLoS Pathogens
Publisher:Public Library of Science
ISSN (Online):1553-7374
Copyright Holders:Copyright © 2018 Kovarova et al.
First Published:First published in PLoS Pathogens 14(3);e1006953
Publisher Policy:Reproduced under a Creative Commons License

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Project CodeAward NoProject NamePrincipal InvestigatorFunder's NameFunder RefLead Dept
371799The Wellcome Centre for Molecular Parasitology ( Core Support )Andrew WatersWellcome Trust (WELLCOTR)104111/Z/14/Z & AIII - PARASITOLOGY
623593Institutional Strategic Support Fund (ISSF)Anna DominiczakWellcome Trust (WELLCOTR)105614/Z/14/ZRI CARDIOVASCULAR & MEDICAL SCIENCES
571301ParaMet - A systematic approach to understanding parasite metabolism.Sylke MullerEuropean Commission (EC)ParaMet290080-FP7III - PARASITOLOGY