Staphylococcus aureus Stress Response to Bicarbonate Depletion
Abstract
:1. Introduction
2. Results
2.1. The mpsABC Deletion Mutant Has a Thicker Cell Wall
2.2. The ΔmpsABC Mutant Is Largely Inert to Cell Lysis Induced by Lysostaphin and Triton X-100
2.3. The Peptidoglycan (PG) of the Mutant Contains Alanine in the Gly5-Bridge
2.4. Comparative Transcriptome Analysis Revealed Differentially Regulated Genes in ΔmpsABC under Ambient Air Condition
2.5. The ΔmpsABC Mutant’s Wall Teichoic Acids (WTA) Showed a Lower α- and Higher ß-Glycosylation
3. Discussion
4. Materials and Methods
4.1. Bacterial Strains and Growth Conditions
4.2. Growth Studies
4.3. RNA Isolation, Library Construction, Sequencing, and Analysis
4.4. qRT-PCR Validation
4.5. Triton X-100-Induced Autolysis Assay
4.6. Lysostaphin-Induced Lysis Assay
4.7. Transmission Electron Microscopy
4.8. Muropeptide Analysis
4.9. Mass Spectrometry Analysis of Monomeric Muropeptides
4.10. Flow Cytometry Analysis with WTA-Detecting Antibodies
4.11. Statistical Analyses and Quantification of CW Thickness
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Cell Wall Thickness in nm (mean ± SD) | ||
---|---|---|
Strain | Ambient Air | 5% CO2 |
JE2 parent | 15.37 ± 2.64 | 12.05 ± 2.44 |
JE2ΔmpsABC | 33.43 ± 7.71 | 15.00 ± 2.78 |
Protein Number | Function | Fold Change Mutant vs. Parent | |
---|---|---|---|
4 h | 8 h | ||
Cell wall (CW) lytic enzymes | |||
B7H15_11645 | SceD, lytic transglycosylase | 592.3 | 517.1 |
B7H15_14290 | IsaA, lytic transglycosylase | 4.3 | 2.1 |
B7H15_12775 | SsaA, secretory antigen | 20 | 5.2 |
B7H15_10875 | Amidase, 251 aa, prophage-encoded | 16.8 | 9.9 |
B7H15_01500 | LytM, glycine-glycine endopeptidase | 14.8 | 8.4 |
B7H15_03720 | LysM, peptidoglycan-binding domain-containing protein, probably autolysin | 8 | 2 |
Wall teichoic acid (WTA) biosynthesis | |||
B7H15_05355 | TarM, poly(ribitol-phosphate) α-N-acetylglucosaminyltransferase | −43.3 | −61.5 |
CW anchored proteins | |||
B7H15_00750 | SasD, cell wall-anchored protein | −27.3 | −3.2 |
B7H15_14775 | SasA, serine-rich repeat glycoprotein adhesin | −23.4 | −15 |
B7H15_13880 | SIRK signal domain/LPXTG anchor domain surface protein | −22.2 | −3.3 |
B7H15_00135 | AdsA, LPXTG-anchored adenosine synthase | −11.5 | −4.1 |
B7H15_00620 | Spa, staphylococcal protein A | −6.4 | −1.6 |
Secreted enzymes | |||
B7H15_01535 | Peptidase C51 domain-containing protein | −64.6 | −31 |
B7H15_14690 | Aureolysin, zinc metalloproteinase | −44.6 | −1.2 |
B7H15_00545 | Phospholipase C, phosphatidylinositol | −23.1 | −1.7 |
B7H15_01765 | Lip2(geh), triacylglycerol lipase | −20.3 | −200 |
Transporters | |||
B7H15_01540 | Type VII secretion effector EsxA (and all other type VII secretion protein genes) | −56.5 | −102 |
B7H15_01675 | Formate/nitrite transporter family protein | −33.7 | −1.7 |
B7H15_01120 | Peptide ABC transporter substrate-binding protein | −15.3 | −4.7 |
B7H15_01110 | ABC transporter permease | −14.9 | −6 |
B7H15_00420 | YdhK family protein, lipoprotein, putative | −13.5 | −29 |
B7H15_00415 | Copper-translocating P-type ATPase | −11.2 | −21.6 |
B7H15_03550 | Metal ABC transporter permease | −11.2 | −176 |
Toxins | |||
B7H15_02365 | PSM-α3, phenol-soluble modulin | −479 | −6360 |
B7H15_11280 | δ-lysin, phenol-soluble modulin | −336 | −11,215 |
B7H15_06120 | PSM-ß, phenol-soluble modulin | −235 | −5200 |
B7H15_06050 | Hla, α-hemolysin | −61 | −2408 |
Regulators | |||
B7H15_14830 | IcaR, ica operon transcriptional repressor | −20.7 | −15.6 |
B7H15_11300 | AgrA, accessory gene regulator protein A | −25 | −91 |
B7H15_11285 | AgrB, accessory gene regulator protein B | −24 | −89 |
B7H15_11290 | AgrD, accessory gene regulator protein D | −24 | −69 |
B7H15_11295 | AgrC, accessory gene regulator protein C | −28 | −90 |
B7H15_03430 | SarA, staphylococcal accessory regulator | −1.7 | −3 |
B7H15_06535 | CodY, transcriptional repressor | −1.2 | −1.6 |
Prophage genes | |||
B7H15_10915 | Phage tail family protein (and most phage-related genes) | 12.5 | nd |
B7H15_11045 | PVL, phi PVL orf 51-like protein | 16.5 | 13 |
Cross bridge synthesis | |||
B7H15_13360 | FmhA, fem-like factor | 1.02 | 1.7 |
B7H15_06500 | FmhC, fem-like factor | 0.98 | −2.3 |
Fatty acid biosynthesis | |||
B7H15_06405 | FabD, ACP S-malonyltransferase | 1.2 | 1.5 |
B7H15_05075 | FabF, ß-ketoacyl-ACP synthase II | −1.5 | −1.2 |
B7H15_06410 | FabG, 3-oxoacyl-acyl-carrier-protein reductase | −1.2 | −1.1 |
B7H15_05210 | FabI, enoyl-ACP reductase | 1.5 | 1.7 |
B7H15_11660 | FabZ, 3-hydroxyacyl-ACP dehydratase | 2.2 | 1.6 |
B7H15_07130 | PlsY, glycerol−3-phosphate acyltransferase | −1.2 | −1.5 |
B7H15_06400 | PlsX, phosphate acyltransferase | −1.5 | 1 |
B7H15_06385 | FakA, fatty acid kinase | 1.1 | 1 |
B7H15_04195 | FakB1, fatty acid binding protein 1 | −1.2 | 1 |
B7H15_07520 | FakB2, fatty acid binding protein 2 | 1 | 1 |
Carboxylating Enzymes | |||
B7H15_09750 | PckA, PEP carboxylase | −5 | −100 |
B7H15_08395 | AccC, acetyl-coA carboxylase | 1.2 | 2 |
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Liberini, E.; Fan, S.-H.; Bayer, A.S.; Beck, C.; Biboy, J.; François, P.; Gray, J.; Hipp, K.; Koch, I.; Peschel, A.; et al. Staphylococcus aureus Stress Response to Bicarbonate Depletion. Int. J. Mol. Sci. 2024, 25, 9251. https://doi.org/10.3390/ijms25179251
Liberini E, Fan S-H, Bayer AS, Beck C, Biboy J, François P, Gray J, Hipp K, Koch I, Peschel A, et al. Staphylococcus aureus Stress Response to Bicarbonate Depletion. International Journal of Molecular Sciences. 2024; 25(17):9251. https://doi.org/10.3390/ijms25179251
Chicago/Turabian StyleLiberini, Elisa, Sook-Ha Fan, Arnold S. Bayer, Christian Beck, Jacob Biboy, Patrice François, Joe Gray, Katharina Hipp, Iris Koch, Andreas Peschel, and et al. 2024. "Staphylococcus aureus Stress Response to Bicarbonate Depletion" International Journal of Molecular Sciences 25, no. 17: 9251. https://doi.org/10.3390/ijms25179251