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A metabolomics approach exploring the function of the ESX-3 type VII secretion system of M. smegmatis

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Abstract

The genome of Mycobacterium, including Mycobacterium tuberculosis, contains five copies of a cluster of genes encoding a novel type VII secretion system, named the ESX gene cluster region. This ESX-3 gene cluster is essential for in vitro growth and is thought to play a role in iron and zinc homeostasis, however, its exact functionality remains an enigma. A metabolomics research approach was subsequently used to compare the metabolite profiles of a M. smegmatis ESX-3 knockout strain to that a wild type parental strain, in order to elucidate its functionality from a metabolic perspective. Statistical analysis of the GC–MS generated data showed a clear separation between the wild type and knockout sample groups, based on the analysed metabolite profiles of these organisms. Of all the metabolite markers identified, various amino acids and metabolite pathways related to these, appeared to be most affected by the ESX-3 knockout, especially those with enzymes regulated by iron and zinc, supporting previous genomics and proteomics generated hypotheses and findings. This study is the first to demonstrate the capacity of using metabolomics, in conjunction with previous genomics and proteomic findings, to identify underlying metabolic changes and confirm previous hypotheses related to the functionality of ESX-3 in Mycobacterium growth and survival.

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Acknowledgments

The authors would like to thank Edith Machowski for generously providing the p2NIL and pGOAL17 vectors used in the construction of the M. smegmatis ESX-3 knockout strain, and Dr Gerhard Koekemoer for assisting in the bioinformatics.

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Authors and Affiliations

  1. School for Physical and Chemical Sciences, Centre for Human Metabonomics, North-West University, Private Bag x6001, Box 269, Potchefstroom, 2531, South Africa

    Du Toit Loots & Reinart J. Meissner-Roloff

  2. DST/NRF Centre of Excellence in Biomedical Tuberculosis Research, US/MRC Centre for Molecular and Cellular Biology, Division of Molecular Biology and Human Genetics, Department of Biomedical Sciences, Faculty of Health Sciences, Stellenbosch University, P.O. Box 19063, Francie van Zijl Drive, Tygerberg, 7505, South Africa

    Mae Newton-Foot & Nicolaas C. Gey van Pittius

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  1. Du Toit Loots
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  2. Reinart J. Meissner-Roloff
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  3. Mae Newton-Foot
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  4. Nicolaas C. Gey van Pittius
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Correspondence to Du Toit Loots.

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Loots, D.T., Meissner-Roloff, R.J., Newton-Foot, M. et al. A metabolomics approach exploring the function of the ESX-3 type VII secretion system of M. smegmatis . Metabolomics 9, 631–641 (2013). https://doi.org/10.1007/s11306-012-0481-x

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  • DOI: https://doi.org/10.1007/s11306-012-0481-x

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