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Mycobacterium smegmatis PrrAB two-component system influences triacylglycerol accumulation during ammonium stress

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  • Authors: Jason D. Maarsingh1​ , Shelley E. Haydel1​,2​,*
    • VIEW AFFILIATIONS
    • 1​ 1​School of Life Sciences, Arizona State University, Tempe, AZ 85287, USA 2​ 2​Biodesign Institute Center for Immunotherapy, Vaccines and Virotherapy, Arizona State University, Tempe, AZ 85287, USA
  • *Correspondence: Shelley E. Haydel [email protected]
  • First Published Online: 07 August 2018, Microbiology 164: 1276-1288, doi: 10.1099/mic.0.000705
  • Subject: Physiology and Metabolism
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Mycobacterium smegmatis PrrAB two-component system influences triacylglycerol accumulation during ammonium stress, Page 1 of 1

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  • The PrrAB two-component system is conserved across all sequenced mycobacterial species and is essential for viability in Mycobacterium tuberculosis, thus making it a promising drug target. The prrAB operon was successfully deleted in nonpathogenic Mycobacterium smegmatis, and the ∆prrAB mutant strain exhibited clumping in ammonium-limited medium and significantly reduced growth during ammonium and hypoxic stress. To assess the influence of M. tuberculosis PrrA overexpression, we constructed a recombinant M. smegmatis ∆prrAB mutant strain which overexpresses M. tuberculosis prrA. M. smegmatis prrAB and M. tuberculosis prrA complemented the M. smegmatis ∆prrAB deletion mutant in Middlebrook M7H9 and ammonium-limited media and during hypoxic and ammonium stress. Based on quantitative untargeted mass spectrometry-based lipidomics, triacylglycerol lipid species were significantly upregulated in the ∆prrAB mutant strain compared to the wild-type when cultured in ammonium-limited medium, revealing that M. smegmatis PrrAB influences triacylglycerol levels during ammonium stress. These results were qualitatively corroborated by thin-layer chromatography. Furthermore, the ∆prrAB mutant significantly upregulated expression of several genes (glpK, GPAT, WS/DGAT, accA3, accD4, accD6 and Ag85C) that participate in triacylglycerol and lipid biosynthetic pathways, thus corroborating the lipidomics analyses.

  • Eight supplementary tables and two supplementary figures are available with the online version of this article.

© 2018 The Authors

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/content/journal/micro/10.1099/mic.0.000705
2018-08-07
2019-06-16
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