Preprint Article Version 1 This version is not peer-reviewed

Staphylococcus aureus Stress Response to Bicarbonate Depletion

Elisa Liberini
,
Sook-Ha Fan
,
Arnold S. Bayer
,
Christian Beck
,
Jacob Biboy
,
Patrice François
,
Joe Gray
,
Katharina Hipp
,
Iris Koch
,
Andreas Peschel
,
Brigitte Sailer
,
Daniela Vollmer
,
Waldemar Vollmer
,
Friedrich Götz
*
Version 1 : Received: 27 July 2024 / Approved: 30 July 2024 / Online: 30 July 2024 (11:03:19 CEST)

How to cite: Liberini, E.; Fan, S.-H.; Bayer, A. S.; Beck, C.; Biboy, J.; François, P.; Gray, J.; Hipp, K.; Koch, I.; Peschel, A.; Sailer, B.; Vollmer, D.; Vollmer, W.; Götz, F. Staphylococcus aureus Stress Response to Bicarbonate Depletion. Preprints 2024, 2024072401. https://doi.org/10.20944/preprints202407.2401.v1 Liberini, E.; Fan, S.-H.; Bayer, A. S.; Beck, C.; Biboy, J.; François, P.; Gray, J.; Hipp, K.; Koch, I.; Peschel, A.; Sailer, B.; Vollmer, D.; Vollmer, W.; Götz, F. Staphylococcus aureus Stress Response to Bicarbonate Depletion. Preprints 2024, 2024072401. https://doi.org/10.20944/preprints202407.2401.v1

Abstract

Bicarbonate and CO2 are essential substrates for carboxylation reactions in bacterial central metabolism. In Staphylococcus aureus, the bicarbonate transporter, MpsABC (membrane potential-generating system) is the only carbon concentrating system. A mpsABC deletion mutant can hardly grow in ambient air. In this study, we investigated the changes that occur in S. aureus when it suffers from CO2/bicarbonate deficiency. Electron microscopy revealed that mpsABC has a twofold thicker cell wall thickness compared to the parent strain. The mutant was also substantially inert to cell lysis induced by lysostaphin and the non-ionic surfactant Triton X-100. Mass spectrometry analysis of muropeptides revealed the incorporation of alanine into the pentaglycine interpeptide bridge, which explains the mutant’s lysostaphin resistance. Flow cytometry analysis of wall teichoic acid (WTA) glycosylation patterns revealed a significantly lower -glycosylated and higher ß-glycosylated WTA, explaining the mutant’s increased resistance towards Triton X-100. Comparative transcriptome analysis showed altered gene expression profiles. Autolysin-encoding genes such as sceD, a lytic transglycosylase encoding gene, were upregulated, while genes related to cell wall-anchored proteins, secreted proteins, transporters, and toxins were downregulated. Overall, we demonstrate that bicarbonate deficiency is a stress response that causes changes in cell wall composition and global gene expression resulting in increased resilience to cell wall lytic enzymes and detergents.

Keywords

Staphylococcus aureus; bicarbonate transporter; MpsABC; cell wall; peptidoglycan; transcriptome

Subject

Biology and Life Sciences, Biochemistry and Molecular Biology

Copyright: This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

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