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Volume 164, Issue 10

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

Abstract

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.

  • Received:
  • Accepted:
  • Published Online:
Keyword(s): lipidomics , Mycobacteria , triacylglycerol and two-component system
© 2018 The Authors
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2018-08-07
2024-04-19
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Mycobacterium smegmatis PrrAB two-component system influences triacylglycerol accumulation during ammonium stress
Microbiology 164, 1276 (2018); https://doi.org/10.1099/mic.0.000705
/content/journal/micro/10.1099/mic.0.000705
/content/journal/micro/10.1099/mic.0.000705
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