Pseudomonas aeruginosa H3-T6SS Combats H2O2 Stress by Diminishing the Amount of Intracellular Unincorporated Iron in a Dps-Dependent Manner and Inhibiting the Synthesis of PQS
Abstract
:1. Introduction
2. Results
2.1. H3-T6SS Inhibits the Synthesis of PQS
2.2. PQS Differentially Regulates the Expression of H3-T6SS Structural Genes and tseF
2.3. H3-T6SS Plays an Important Role in Combating Oxidative Stress Caused by H2O2
2.4. H3-T6SS Is Positively Regulated by RpoS
2.5. H3-T6SS Affects the Expression of Various Proteins Related to Oxidative Stress
2.6. P. aeruginosa H3-T6SS Combats H2O2 Stress in Two Ways
2.6.1. H3-T6SS Combats H2O2 Stress by Reducing Intracellular Free Fe2+
2.6.2. H3-T6SS Combats H2O2 Stress by Inhibiting the Synthesis of PQS
3. Discussion
4. Materials and Methods
4.1. Bacterial Strains and Growth Conditions
4.2. Plasmid Construction
4.3. Construction of Chromosomal Fusion Reporter Strains
4.4. β-Galactosidase Assays
4.5. Liquid Chromatography-Mass Spectrometry Analysis
4.6. Sensitivity Assays
4.7. Intracellular ROS Detection
4.8. Proteomic Analysis
4.9. Electron Paramagnetic Resonance Spectroscopy Analysis
4.10. Statistical Analysis
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Conflicts of Interest
References
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PA No. a | Gene | Fold Change b | p-Values | Protein Description c |
---|---|---|---|---|
PQS biosynthesis | ||||
PA0999 | pqsD | 29.6 | 2.23 × 10−7 | 3-oxoacyl-ACP synthase |
PA2080 | kynU | 10.9 | 7.36 × 10−3 | Kynureninase KynU |
Oxidative processes and oxidative stress | ||||
PA0140 | ahpF | 10.9 | 7.36 × 10−3 | Alkyl hydroperoxide reductase |
PA3529 | 10.9 | 7.36 × 10−3 | Peroxidase | |
PA5240 | trxA | 3.4 | 8.51 × 10−4 | Thioredoxin |
General stress response | ||||
PA4385 | groEL | 15.6 | 5.92 × 10−4 | Molecular chaperone GroEL |
PA4386 | groES | 29.6 | 2.23 × 10−7 | Co-chaperonin GroES |
PA4761 | dnaK | 2.3 | 1.68 × 10−3 | Molecular chaperone DnaK |
PA0962 | dps | −21.8 | 8.05 × 10−7 | DNA-binding stress protein |
Genes of the oxidative stress response d | ||||
PA0102 | 15.6 | 5.92 × 10−4 | Carbonic anhydrase | |
PA5015 | aceE | 3.0 | 5.12 × 10−5 | Pyruvate dehydrogenase subunit E1 |
PA1973 | pqqF | 15.6 | 5.92 × 10−4 | Pyrroloquinoline quinone biosynthesis protein F |
PA4131 | 8.6 | 2.44 × 10−6 | Iron-sulfur protein | |
PA3820 | secF | 15.6 | 5.92 × 10−4 | Preprotein translocase subunit SecF |
PA3821 | secD | 5.1 | 2.19 × 10−3 | Preprotein translocase subunit SecD |
PA5128 | secB | 10.9 | 7.36 × 10−3 | Preprotein translocase subunit SecB |
PA1583 | sdhA | −2.3 | 4.72 × 10−2 | Succinate dehydrogenase flavoprotein subunit |
PA1880 | −2.6 | 2.32 × 10−2 | Oxidoreductase | |
PA4694 | ilvC | −3.9 | 7.04 × 10−6 | Ketol-acid reductoisomerase |
Iron sequestration | ||||
PA3397 | fpr | 5.1 | 2.19 × 10−3 | Ferredoxin-NADP reductase |
PA4235 | bfrA | −1.6 | 2.01 × 10−2 | Bacterioferritin |
PA0962 | dps | −21.8 | 8.05 × 10−7 | DNA-binding stress protein |
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Lin, J.; Yang, J.; Cheng, J.; Zhang, W.; Yang, X.; Ding, W.; Zhang, H.; Wang, Y.; Shen, X. Pseudomonas aeruginosa H3-T6SS Combats H2O2 Stress by Diminishing the Amount of Intracellular Unincorporated Iron in a Dps-Dependent Manner and Inhibiting the Synthesis of PQS. Int. J. Mol. Sci. 2023, 24, 1614. https://doi.org/10.3390/ijms24021614
Lin J, Yang J, Cheng J, Zhang W, Yang X, Ding W, Zhang H, Wang Y, Shen X. Pseudomonas aeruginosa H3-T6SS Combats H2O2 Stress by Diminishing the Amount of Intracellular Unincorporated Iron in a Dps-Dependent Manner and Inhibiting the Synthesis of PQS. International Journal of Molecular Sciences. 2023; 24(2):1614. https://doi.org/10.3390/ijms24021614
Chicago/Turabian StyleLin, Jinshui, Jianshe Yang, Juanli Cheng, Weipeng Zhang, Xu Yang, Wei Ding, Heng Zhang, Yao Wang, and Xihui Shen. 2023. "Pseudomonas aeruginosa H3-T6SS Combats H2O2 Stress by Diminishing the Amount of Intracellular Unincorporated Iron in a Dps-Dependent Manner and Inhibiting the Synthesis of PQS" International Journal of Molecular Sciences 24, no. 2: 1614. https://doi.org/10.3390/ijms24021614