An essential bifunctional enzyme in Mycobacterium tuberculosis for itaconate dissimilation and leucine catabolism

Wang, H. , Federov, A. A., Federov, E. V., Hunt, D. M., Rodgers, A., Douglas, H. L., Garza-Garcia, A., Bonanno, J. B., Almo, S. C. and Sório de Carvalho, L. P. (2019) An essential bifunctional enzyme in Mycobacterium tuberculosis for itaconate dissimilation and leucine catabolism. Proceedings of the National Academy of Sciences of the United States of America, 116(32), pp. 15907-15913. (doi: 10.1073/pnas.1906606116) (PMID:31320588) (PMCID:PMC6689899)

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Abstract

Mycobacterium tuberculosis (Mtb) is the etiological agent of tuberculosis. One-fourth of the global population is estimated to be infected with Mtb, accounting for ∼1.3 million deaths in 2017. As part of the immune response to Mtb infection, macrophages produce metabolites with the purpose of inhibiting or killing the bacterial cell. Itaconate is an abundant host metabolite thought to be both an antimicrobial agent and a modulator of the host inflammatory response. However, the exact mode of action of itaconate remains unclear. Here, we show that Mtb has an itaconate dissimilation pathway and that the last enzyme in this pathway, Rv2498c, also participates in L-leucine catabolism. Our results from phylogenetic analysis, in vitro enzymatic assays, X-ray crystallography, and in vivo Mtb experiments, identified Mtb Rv2498c as a bifunctional β-hydroxyacyl-CoA lyase and that deletion of the rv2498c gene from the Mtb genome resulted in attenuation in a mouse infection model. Altogether, this report describes an itaconate resistance mechanism in Mtb and an L-leucine catabolic pathway that proceeds via an unprecedented (R)-3-hydroxy-3-methylglutaryl-CoA (HMG-CoA) stereospecific route in nature.

Item Type:Articles
Additional Information:We thank Dr. Geoff Kelly (Medical Research Council [MRC] Biomedical NMR Centre) for assistance with NMR spectroscopy. NMR data were recorded in the MRC Biomedical NMR Centre at the Francis Crick Institute, which receives core funding from Cancer Research UK Grant FC001029; Medical Research Council Grant FC001029; and Wellcome Trust Grant FC001029. Work in L.P.S.d.C.’s laboratory was supported by the Francis Crick Institute, which receives its core funding from Cancer Research UK Grant FC001060, UK MRC Grant FC001060, and Wellcome Trust Grant FC001060. L.P.S.d.C.’s laboratory also acknowledges funds from Wellcome Trust New Investigator Award 104785/B/14/Z. This research used resources of the Advanced Photon Source, a US Department of Energy (DOE) Office of Science User Facility operated for the DOE Office of Science by Argonne National Laboratory under Contract DE-AC02-06CH11357. Use of the Lilly Research Laboratories Collaborative Access Team beamline at Sector 31 of the Advanced Photon Source was provided by Eli Lilly Company, which operates the facility. The Einstein Crystallographic Core X-Ray diffraction facility is supported by NIH Shared Instrumentation Grant S10 OD020068, which we gratefully acknowledge. S.C.A. acknowledges support from the US NIH Grant P01 GM118303.
Status:Published
Refereed:Yes
Glasgow Author(s) Enlighten ID:Wang, Dr Hua
Authors: Wang, H., Federov, A. A., Federov, E. V., Hunt, D. M., Rodgers, A., Douglas, H. L., Garza-Garcia, A., Bonanno, J. B., Almo, S. C., and Sório de Carvalho, L. P.
College/School:College of Medical Veterinary and Life Sciences > Institute of Infection Immunity and Inflammation
Journal Name:Proceedings of the National Academy of Sciences of the United States of America
Publisher:National Academy of Sciences
ISSN:0027-8424
ISSN (Online):1091-6490
Published Online:18 July 2019
Copyright Holders:Copyright © 2019 The Authors
First Published:First published in Proceedings of the National Academy of Sciences of the United States of America 116(32):15907-15913
Publisher Policy:Reproduced under a Creative Commons License

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