Bisphenol S Promotes the Transfer of Antibiotic Resistance Genes via Transformation
<p>BPS promotes the transformation of ARGs into <span class="html-italic">E. coli</span> DH5α. (<b>A</b>) Growth curves of the recipient bacterium (<span class="html-italic">E. coli</span> DH5α) in the presence of different concentrations of the BPS (0.1–10 μg/mL). (<b>B</b>) Effects of different concentrations of the BPS on the frequency of transformation of pUC19 plasmid into <span class="html-italic">E. coli</span> DH5α. Statistically significant differences were determined using one-way ANOVA at * <span class="html-italic">p</span> < 0.05, ** <span class="html-italic">p</span> < 0.01 and *** <span class="html-italic">p</span> < 0.001, respectively. NS, not significant. (<b>C</b>) Gel electropherograms of pUC19 plasmid, recipient bacteria, and transformants at different concentrations of BPS. (<b>D</b>) MIC values of recipient bacteria and transformants.</p> "> Figure 2
<p>BPS stimulates the production of ROS and enhances membrane permeability in the recipient bacteria. (<b>A</b>) Effects of different concentrations of BPS on ROS production by recipient bacteria. (<b>B</b>) Heat map of increased expression levels of genes related to the oxidative stress system and SOS response system of bacteria after BPS treatment. (<b>C</b>) Changes in outer membrane permeability of recipient bacteria following BPS pressure. (<b>D</b>) Changes in inner membrane permeability in response to BPS treatment. (<b>E</b>) Effect of BPS on membrane fluidity. Statistically significant differences were determined using one-way ANOVA at * <span class="html-italic">p</span> < 0.05, ** <span class="html-italic">p</span> < 0.01, *** <span class="html-italic">p</span> < 0.001 and **** <span class="html-italic">p</span> < 0.0001, respectively. NS, not significant. (<b>F</b>) Heatmap of the increased expression levels of genes related to bacterial membrane permeability after BPS treatment. (<b>G</b>) SEM images of <span class="html-italic">E. coli</span> DH5α bacterial cells exposed to 0.5 μg/mL BPS for 4 h. Cell membrane damage is indicated by red arrows.</p> "> Figure 3
<p>BPS enhances bacterial metabolism by accelerating the TCA cycle. (<b>A</b>) Bacterial respiration levels of <span class="html-italic">E. coli</span> DH5α were unchanged or even decreased under the pressure of BPS. (<b>B</b>) Heatmap of the expression levels of genes related to bacterial electron transport chain in response to BPS treatment. (<b>C</b>) Heatmap of TCA cycle-related gene expression levels in response to BPS. Bacterial (<b>D</b>) NAD<sup>+</sup>/NADH ratio, (<b>E</b>) NAD<sup>+</sup> content, and (<b>F</b>) NADH content under BPS treatment. Statistically significant differences were determined using one-way ANOVA at ** <span class="html-italic">p</span> < 0.01, *** <span class="html-italic">p</span> < 0.001 and **** <span class="html-italic">p</span> < 0.0001, respectively. NS, not significant.</p> "> Figure 4
<p>BPS stimulates ATP synthesis and flagellar motility. (<b>A</b>) ΔpH changes of recipient bacteria in response to BPS treatment, measured using BCECF. (<b>B</b>) Membrane potential of recipient bacteria in response to BPS stress, monitored using DiSC<sub>3</sub>(5). (<b>C</b>) Bacterial ATP synthesis after exposure to BPS. (<b>D</b>) Heat map of the expression level of bacterial ATP synthase-related genes under BPS stress. (<b>E</b>) Heatmap of the expression level of bacterial flagellum-related genes after BPS treatment. (<b>F</b>) Swimming motility test of <span class="html-italic">E. coli</span> DH5α under BPS stress, scale bar, 0.5 cm. Statistically significant differences were determined using one-way ANOVA at **** <span class="html-italic">p</span> < 0.0001. NS, not significant.</p> "> Figure 5
<p>Schematic diagram of the mechanism of increased transformation by BPS treatment. The frequency of transformation of antibiotic-resistant plasmids was significantly increased under the stress of low concentrations of BPS. Potential mechanisms include a dramatic increase in ROS production and activation of the SOS response, which increases membrane permeability and fluidity. In addition, the accelerated TCA cycle generates a large amount of ATP, and flagellar motility was also enhanced. These actions are favorable for plasmid uptake, facilitation, and integration.</p> ">
Abstract
:1. Introduction
2. Results
2.1. BPS Promotes the Spread of ARGs via Transformation
2.2. BPS Enhances Bacterial Oxidative Stress and Activates SOS Response
2.3. BPS Improves Membrane Permeability and Fluidity
2.4. BPS Enhances Metabolic States by Accelerating TCA Cycle
2.5. BPS Stimulates ATP Supply and Improves Flagellar Motility
3. Discussion
4. Materials and Methods
4.1. Bacterial Strains, Plasmids, Antibiotics, BPS, and Culture Media
4.2. Measurement of Growth Curves
4.3. Determination of Minimum Inhibitory Concentration (MIC)
4.4. Establishment of Transformation System and Validation of Transformants
4.5. Detection of Oxidative Stress Levels in Bacteria
4.6. Detection of Bacterial Membrane Permeability and Fluidity
4.7. Scanning Electron Microscope (SEM) Analysis
4.8. Detection of Bacterial NAD+/NADH
4.9. Detection of Bacterial Proton Motive Force (PMF)
4.10. Assay of Bacterial Motility
4.11. Monitoring of Bacterial Respiration Levels
4.12. Content Determination of Bacterial ATP
4.13. RNA Extraction and RT-qPCR Analysis
4.14. Statistical Analysis
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Conflicts of Interest
References
- Darby, E.M.; Trampari, E.; Siasat, P.; Gaya, M.S.; Alav, I.; Webber, M.A.; Blair, J.M.A. Molecular mechanisms of antibiotic resistance revisited. Nat. Rev. Microbiol. 2022, 21, 280–295. [Google Scholar] [CrossRef] [PubMed]
- Bhardwaj, S.; Mehra, P.; Dhanjal, D.S.; Sharma, P.; Sharma, V.; Singh, R.; Nepovimova, E.; Chopra, C.; Kuca, K. Antibiotics and Antibiotic Resistance-Flipsides of the Same Coin. Curr. Pharm. Des. 2022, 28, 2312–2329. [Google Scholar] [CrossRef] [PubMed]
- Kohanski, M.A.; DePristo, M.A.; Collins, J.J. Sublethal antibiotic treatment leads to multidrug resistance via radical-induced mutagenesis. Mol. Cell 2010, 37, 311–320. [Google Scholar] [CrossRef] [PubMed]
- Lopatkin, A.J.; Huang, S.; Smith, R.P.; Srimani, J.K.; Sysoeva, T.A.; Bewick, S.; Karig, D.K.; You, L. Antibiotics as a selective driver for conjugation dynamics. Nat. Microbiol. 2016, 1, 16044. [Google Scholar] [CrossRef] [PubMed]
- Yu, Z.; Wang, Y.; Henderson, I.R.; Guo, J. Artificial sweeteners stimulate horizontal transfer of extracellular antibiotic resistance genes through natural transformation. ISME J. 2022, 16, 543–554. [Google Scholar] [CrossRef]
- Liao, J.; Chen, Y.; Huang, H. Effects of CO2 on the transformation of antibiotic resistance genes via increasing cell membrane channels. Environ. Pollut. 2019, 254, 113045. [Google Scholar] [CrossRef]
- Zhu, S.; Yang, B.; Jia, Y.; Yu, F.; Wang, Z.; Liu, Y. Comprehensive analysis of disinfectants on the horizontal transfer of antibiotic resistance genes. J. Hazard. Mater. 2023, 453, 131428. [Google Scholar] [CrossRef]
- Weatherly, L.M.; Gosse, J.A. Triclosan exposure, transformation, and human health effects. J. Toxicol. Environ. Health B Crit. Rev. 2017, 20, 447–469. [Google Scholar] [CrossRef]
- Ma, Y.; Liu, H.; Wu, J.; Yuan, L.; Wang, Y.; Du, X.; Wang, R.; Marwa, P.W.; Petlulu, P.; Chen, X.; et al. The adverse health effects of bisphenol A and related toxicity mechanisms. Environ. Res. 2019, 176, 108575. [Google Scholar] [CrossRef]
- Manzan-Martins, C.; Paulesu, L. Impact of bisphenol A (BPA) on cells and tissues at the human materno-fetal interface. Tissue Cell 2021, 73, 101662. [Google Scholar] [CrossRef]
- Santoro, A.; Chianese, R.; Troisi, J.; Richards, S.; Nori, S.L.; Fasano, S.; Guida, M.; Plunk, E.; Viggiano, A.; Pierantoni, R.; et al. Neuro-toxic and Reproductive Effects of BPA. Curr. Neuropharmacol. 2019, 17, 1109–1132. [Google Scholar] [CrossRef] [PubMed]
- Lao, X.; Tam, N.F.Y.; Zhong, M.; Wu, Q.; Liu, Z.; Huang, X.; Wei, L.; Liu, Y.; Luo, D.; Li, S. Distribution and risk assessment of antibiotic and bisphenol compounds residues in drinking water sources of Guangdong. Environ. Earth Sci. 2024, 83, 475. [Google Scholar] [CrossRef]
- Zhang, J.; Yao, K.; Yin, J.; Lyu, B.; Zhao, Y.; Li, J.; Shao, B.; Wu, Y. Exposure to Bisphenolic Analogues in the Sixth Total Diet Study—China, 2016–2019. China CDC Wkly. 2022, 4, 180–184. [Google Scholar] [CrossRef] [PubMed]
- Gingrich, J.; Pu, Y.; Roberts, J.; Karthikraj, R.; Kannan, K.; Ehrhardt, R.; Veiga-Lopez, A. Gestational bisphenol S impairs placental endocrine function and the fusogenic trophoblast signaling pathway. Arch. Toxicol. 2018, 92, 1861–1876. [Google Scholar] [CrossRef]
- Sreedevi, P.R.; Suresh, K. Cold atmospheric plasma mediated cell membrane permeation and gene delivery-empirical interventions and pertinence. Adv. Colloid Interface Sci. 2023, 320, 102989. [Google Scholar] [CrossRef]
- Cao, P.; Wall, D. The Fluidity of the Bacterial Outer Membrane Is Species Specific: Bacterial Lifestyles and the Emergence of a Fluid Outer Membrane. Bioessays 2020, 42, e1900246. [Google Scholar] [CrossRef]
- Le, D.; Krasnopeeva, E.; Sinjab, F.; Pilizota, T.; Kim, M. Active Efflux Leads to Heterogeneous Dissipation of Proton Motive Force by Protonophores in Bacteria. mBio 2021, 12, e0067621. [Google Scholar] [CrossRef]
- You, Y.; Ye, F.; Mao, W.; Yang, H.; Lai, J.; Deng, S. An overview of the structure and function of the flagellar hook FlgE protein. World J. Microbiol. Biotechnol. 2023, 39, 126. [Google Scholar] [CrossRef]
- Arsene, M.M.J.; Davares, A.K.L.; Viktorovna, P.I.; Andreevna, S.L.; Sarra, S.; Khelifi, I.; Sergueievna, D.M. The public health issue of antibiotic residues in food and feed: Causes, consequences, and potential solutions. Vet. World 2022, 15, 662–671. [Google Scholar] [CrossRef]
- Yang, Y.; Yang, X.; Zhou, H.; Niu, Y.; Li, J.; Fu, X.; Wang, S.; Xue, B.; Li, C.; Zhao, C.; et al. Bisphenols Promote the Pheromone-Responsive Plasmid-Mediated Conjugative Transfer of Antibiotic Resistance Genes in Enterococcus faecalis. Environ. Sci. Technol. 2022, 56, 17653–17662. [Google Scholar] [CrossRef]
- Wu, L.H.; Zhang, X.M.; Wang, F.; Gao, C.J.; Chen, D.; Palumbo, J.R.; Guo, Y.; Zeng, E.Y. Occurrence of bisphenol S in the environment and implications for human exposure: A short review. Sci. Total Environ. 2018, 615, 87–98. [Google Scholar] [CrossRef] [PubMed]
- Chen, D.; Kannan, K.; Tan, H.L.; Zheng, Z.G.; Feng, Y.L.; Wu, Y.; Widelka, M. Bisphenol Analogues Other Than BPA: Environmental Occurrence, Human Exposure, and Toxicity-A Review. Environ. Sci. Technol. 2016, 50, 5438–5453. [Google Scholar] [CrossRef] [PubMed]
- Zhang, S.; Wang, Y.; Lu, J.; Yu, Z.; Song, H.; Bond, P.L.; Guo, J. Chlorine disinfection facilitates natural transformation through ROS-mediated oxidative stress. ISME J. 2021, 15, 2969–2985. [Google Scholar] [CrossRef] [PubMed]
- Johnston, C.; Martin, B.; Fichant, G.; Polard, P.; Claverys, J.P. Bacterial transformation: Distribution, shared mechanisms and divergent control. Nat. Rev. Microbiol. 2014, 12, 181–196. [Google Scholar] [CrossRef] [PubMed]
- Ma, L.N.; Mi, H.F.; Xue, Y.X.; Wang, D.; Zhao, X.L. The mechanism of ROS regulation of antibiotic resistance and antimicrobial lethality. Yi Chuan 2016, 38, 902–909. [Google Scholar]
- Zhang, S.; Yang, M.J.; Peng, B.; Peng, X.X.; Li, H. Reduced ROS-mediated antibiotic resistance and its reverting by glucose in Vibrio alginolyticus. Environ. Microbiol. 2020, 22, 4367–4380. [Google Scholar] [CrossRef]
- Mantilla-Calderon, D.; Plewa, M.J.; Michoud, G.; Fodelianakis, S.; Daffonchio, D.; Hong, P.Y. Water Disinfection Byproducts Increase Natural Transformation Rates of Environmental DNA in Acinetobacter baylyi ADP1. Environ. Sci. Technol. 2019, 53, 6520–6528. [Google Scholar] [CrossRef]
- Zhang, Y.; Gu, A.Z.; He, M.; Li, D.; Chen, J. Subinhibitory concentrations of disinfectants promote the horizontal transfer of multidrug resistance genes within and across genera. Environ. Sci. Technol. 2017, 51, 570–580. [Google Scholar] [CrossRef]
- Yu, Z.; Wang, Y.; Lu, J.; Bond, P.L.; Guo, J. Nonnutritive sweeteners can promote the dissemination of antibiotic resistance through conjugative gene transfer. ISME J. 2021, 15, 2117–2130. [Google Scholar] [CrossRef]
- Simon, S.M.; Sousa, F.J.; Mohana-Borges, R.; Walker, G.C. Regulation of Escherichia coli SOS mutagenesis by dimeric intrinsically disordered umuD gene products. Proc. Natl. Acad. Sci. USA 2008, 105, 1152–1157. [Google Scholar] [CrossRef]
- Kulbacka, J.; Choromanska, A.; Rossowska, J.; Wezgowiec, J.; Saczko, J.; Rols, M.P. Cell Membrane Transport Mechanisms: Ion Channels and Electrical Properties of Cell Membranes. Adv. Anat. Embryol. Cell Biol. 2017, 227, 39–58. [Google Scholar] [PubMed]
- Lu, J.; Wang, Y.; Zhang, S.; Bond, P.; Yuan, Z.; Guo, J. Triclosan at environmental concentrations can enhance the spread of extracellular antibiotic resistance genes through transformation. Sci. Total Environ. 2020, 713, 136621. [Google Scholar] [CrossRef]
- Henry, R.; Crane, B.; Powell, D.; Deveson Lucas, D.; Li, Z.; Aranda, J.; Harrison, P.; Nation, R.L.; Adler, B.; Harper, M.; et al. The transcriptomic response of Acinetobacter baumannii to colistin and doripenem alone and in combination in an in vitro pharmacokinetics/pharmacodynamics model. J. Antimicrob. Chemother. 2015, 70, 1303–1313. [Google Scholar] [CrossRef] [PubMed]
- Zhou, Y.; Yong, Y.; Zhu, C.; Yang, H.; Fang, B. Exogenous D-ribose promotes gentamicin treatment of several drug-resistant Salmonella. Front. Microbiol. 2022, 13, 1053330. [Google Scholar] [CrossRef] [PubMed]
- Judge, A.; Dodd, M.S. Metabolism. Essays Biochem. 2020, 64, 607–647. [Google Scholar] [CrossRef]
- Alvarez-Rodriguez, I.; Arana, L.; Ugarte-Uribe, B.; Gomez-Rubio, E.; Martin-Santamaria, S.; Garbisu, C.; Alkorta, I. Type IV Coupling Proteins as Potential Targets to Control the Dissemination of Antibiotic Resistance. Front. Mol. Biosci. 2020, 7, 201. [Google Scholar] [CrossRef] [PubMed]
- Martin, I.V.; MacNeill, S.A. ATP-dependent DNA ligases. Genome Biol. 2002, 3, REVIEWS3005. [Google Scholar] [CrossRef]
- Mehring, A.; Erdmann, N.; Walther, J.; Stiefelmaier, J.; Strieth, D.; Ulber, R. A simple and low-cost resazurin assay for vitality assessment across species. J. Biotechnol. 2021, 333, 63–66. [Google Scholar] [CrossRef]
Disclaimer/Publisher’s Note: The statements, opinions and data contained in all publications are solely those of the individual author(s) and contributor(s) and not of MDPI and/or the editor(s). MDPI and/or the editor(s) disclaim responsibility for any injury to people or property resulting from any ideas, methods, instructions or products referred to in the content. |
© 2024 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
Share and Cite
Zhang, J.; Zhu, S.; Sun, J.; Liu, Y. Bisphenol S Promotes the Transfer of Antibiotic Resistance Genes via Transformation. Int. J. Mol. Sci. 2024, 25, 9819. https://doi.org/10.3390/ijms25189819
Zhang J, Zhu S, Sun J, Liu Y. Bisphenol S Promotes the Transfer of Antibiotic Resistance Genes via Transformation. International Journal of Molecular Sciences. 2024; 25(18):9819. https://doi.org/10.3390/ijms25189819
Chicago/Turabian StyleZhang, Jiayi, Shuyao Zhu, Jingyi Sun, and Yuan Liu. 2024. "Bisphenol S Promotes the Transfer of Antibiotic Resistance Genes via Transformation" International Journal of Molecular Sciences 25, no. 18: 9819. https://doi.org/10.3390/ijms25189819