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Metabolic profile of Mycobacterium smegmatis reveals Mce4 proteins are relevant for cell wall lipid homeostasis

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Abstract

Introduction

The Mce proteins are encoded in a variable number of operons (from one to eight) in all Mycobacterium species. A role in the transport of host and mycobacterial lipids has been demonstrated for some Mce proteins in the pathogen Mycobacterium tuberculosis but little is known about these proteins in Mycobacterium smegmatis, a soil dweller species.

Objective

To investigate the role of Mce proteins in M. smegmatis.

Method

We used a non-targeted GC–MS approach to define the metabolic profiles of knockout mutants in mce operons of M. smegmatis. Metabolomic analysis was complemented with thin layer chromatography and transcriptional studies.

Result

We demonstrated that the lack of Mce4 proteins alters the primary carbon metabolism of M. smegmatis by reducing the content of fatty acids and increasing the accumulation of two stress-related compounds, glutamate/glutamine and triacyl glycerol (TAG). We also provide evidence supporting that the accumulation of TAG in a Δmce4 mutant depends on the bacterial redox state.

Conclusion

These results, together with the finding that the cell surface of the Δmce4 mutant is significantly altered, support a role for Mce4 in maintaining the cell wall lipids architecture of M. smegmatis, which, when altered, induces a redox imbalance that results in the accumulation of the stress-related compounds.

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Acknowledgments

This work was funded by INTA Grant PNBIO1131034 and National Institutes of Health (NIH) Grants 1 R01 AI083084 and PAR08-130. MPS, FB, LK, MF and FB are CONICET fellows.

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Author notes

  1. Adam Heuberger

    Present address: Department of Horticulture & Landscape Architecture, Colorado State University, 209 Shepardson Building, Fort Collins, CO, 80523-1173, USA

Authors and Affiliations

  1. CICVyA. Instituto Nacional de Tecnología Agropecuaria, CONICET, B1686IGC, Hurlingham, Buenos Aires, Argentina

    María Paz Santangelo, Federico Blanco, Marina Forrellad, Laura Klepp & Fabiana Bigi

  2. CICVyA, Instituto Nacional de Tecnología Agropecuaria, B1686IGC, Hurlingham, Buenos Aires, Argentina

    Catalina Taibo & Julia Sabio García

  3. Proteomics and Metabolomics Facility, Colorado State University, Microbiology Building, Fort Collins, CO, 80523-2021, USA

    Adam Heuberger

  4. Systems Biology Program, Centro Nacional de Biotecnología, CSIC. C/Darwin 3, Campus de Cantoblanco, 28049, Madrid, Spain

    Pablo I. Nikel

  5. Department of Microbiology Immunology and Pathology College of Veterinary Medicine and Biomedical Sciences, Colorado State University, Fort Collins, CO, 80523-1682, USA

    Mary Jackson

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  1. María Paz Santangelo
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Correspondence to Fabiana Bigi.

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María Paz, Santangelo; Adam, Heuberger; Federico, Blanco; Marina, Forrellad; Catalina, Taibo; Laura, Klepp; Julia, Sabio García; Pablo I., Nikel; Mary, Jackson and Fabiana, Bigi declare that they have no conflict of interest.

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Santangelo, M.P., Heuberger, A., Blanco, F. et al. Metabolic profile of Mycobacterium smegmatis reveals Mce4 proteins are relevant for cell wall lipid homeostasis. Metabolomics 12, 97 (2016). https://doi.org/10.1007/s11306-016-1035-4

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