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Mycobacterium smegmatis PrrAB two-component system influences triacylglycerol accumulation during ammonium stress, Page 1 of 1
< Previous page | Next page > /docserver/preview/fulltext/micro/164/10/1276_micro000705-1.gifThe 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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