Metabolic reprogramming by glutathione S-transferase enhances environmental adaptation of Streptococcus mutans

doi:10.7518/hxkq.2025.2025084

Abstract

Abstract:

Objective This study aims to investigate the impact of glutathione S-transferase (GST) on the environmental adaptability of Streptococcus mutans (S. mutans). Methods A GST knockout strain ΔgsT was constructed. Transcriptomic sequencing was performed to analyze the gene expression differences between the wild-type S. mutans UA159 and its GST knockout strain ΔgsT. Comprehensive functional assessments, including acid tolerance assays, hydrogen peroxide challenge assays, nutrient limitation growth assays, and fluorescence in situ hybridization, were conducted to evaluate the acid tolerance, antioxidant stress resistance, growth kinetics, and interspecies competitive ability of ΔgsT within plaque biofilms. Results Compared with the wild-type S. mutans, 198 genes in ΔgsT were significantly differentially expressed and enriched in pathways related to metabolism, stress response, and energy homeostasis. The survival rate of ΔgsT in acid tolerance assays was markedly reduced (P<0.01). After 15 min of hydrogen peroxide challenge, the survival rate of ΔgsT decreased to 38.12% (wild type, 71.75%). Under nutrient-limiting conditions, ΔgsT exhibited a significantly lower final OD₆₀₀ value than the wild-type strain (P<0.05). In the biofilm competition assays, the proportion of S. mutans ΔgsT in the mixed biofilm (8.50%) was significantly lower than that of the wild type (16.89%) (P<0.05). Conclusion GST enhances the acid resistance, oxidative stress tolerance, and nutrient adaptation of S. mutans by regulating metabolism-related and stress response-related genes.

Key words: glutathione S-transferase, Streptococcus mutans, acid tolerance, oxidative stress, interspecies competition, gene expression regulation, metabolic reprogramming

CLC Number:

R780.2

Zheng Haoyue, Peng Xian, Zou Jing. Metabolic reprogramming by glutathione S-transferase enhances environmental adaptation of Streptococcus mutans[J]. West China Journal of Stomatology, 2025, 43(5): 728-735.

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References 28

[1]	Banas JA. Virulence properties of Streptococcus mutans [J]. Front Biosci, 2004, 9: 1267-1277.
[2]	Featherstone JD. Dental caries: a dynamic disease process[J]. Aust Dent J, 2008, 53(3): 286-291.
[3]	Xu YF, Itzek A, Kreth J. Comparison of genes required for H₂O₂ resistance in Streptococcus gordonii and Streptococcus sanguinis [J]. Microbiology, 2014, 160(12): 2627-2638.
[4]	Sherrill C, Fahey RC. Import and metabolism of glutathione by Streptococcus mutans [J]. J Bacteriol, 1998, 180(6): 1454-1459.
[5]	Vergauwen B, Verstraete K, Senadheera DB, et al. Mole-cular and structural basis of glutathione import in Gram-positive bacteria via GshT and the cystine ABC impor-ter TcyBC of Streptococcus mutans [J]. Mol Microbiol, 2013, 89(2): 288-303.
[6]	Zhang J, Ye ZW, Singh S, et al. An evolving understan-ding of the S-glutathionylation cycle in pathways of redox regulation[J]. Free Radic Biol Med, 2018, 120: 204-216.
[7]	Mailloux RJ. Protein S-glutathionylation reactions as a global inhibitor of cell metabolism for the desensitization of hydrogen peroxide signals[J]. Redox Biol, 2020, 32: 101472.
[8]	Bischoff ME, Shamsaei B, Yang JC, et al. Copper drives remodeling of metabolic state and progression of clear cell renal cell carcinoma[J]. Cancer Discov, 2025, 15(2): 401-426.
[9]	Romig M, Eberwein M, Deobald D, et al. Reactivation and long-term stabilization of the [NiFe] Hox hydrogenase of Synechocystis sp. PCC6803 by glutathione after oxygen exposure[J]. J Biol Chem, 2025, 301(1): 108086.
[10]	Li ZY, Zhang CZ, Li C, et al. S-glutathionylation proteome profiling reveals a crucial role of a thioredoxin-like protein in interspecies competition and cariogene-city of Streptococcus mutans [J]. PLoS Pathog, 2020, 16(7): e1008774.
[11]	Hayes JD, Flanagan JU, Jowsey IR. Glutathione transfe-rases[J]. Annu Rev Pharmacol Toxicol, 2005, 45: 51-88.
[12]	Armstrong RN. Structure, catalytic mechanism, and evolution of the glutathione transferases[J]. Chem Res Toxicol, 1997, 10(1): 2-18.
[13]	Zhang SS, Zhang C, Sun FJ, et al. Glutathione-S-transferase (GST) catalyzes the degradation of chlorimuron-ethyl by Klebsiella jilinsis 2N3[J]. Sci Total Environ, 2020, 729: 139075.
[14]	Kammerscheit X, Chauvat F, Cassier-Chauvat C. From cyanobacteria to human, MAPEG-type glutathione-S-transferases operate in cell tolerance to heat, cold, and lipid peroxidation[J]. Front Microbiol, 2019, 10: 2248.
[15]	Peng X, Zhou XD, Xu X. The oral microbiome bank of China[J]. Int J Oral Sci, 2018, 10(2):16.
[16]	Jones AS. The isolation of bacterial nucleic acids using cetyltrimethylammonium bromide (cetavlon)[J]. Biochim Biophys Acta, 1953, 10(4): 607-612.
[17]	Belli WA, Marquis RE. Adaptation of Streptococcus mutans and Enterococcus hirae to acid stress in continuous culture[J]. Appl Environ Microbiol, 1991, 57(4): 1134-1138.
[18]	Wen ZT, Suntharaligham P, Cvitkovitch DG, et al. Trigger factor in Streptococcus mutans is involved in stress tolerance, competence development, and biofilm formation[J]. Infect Immun, 2005, 73(1): 219-225.
[19]	Klug B, Rodler C, Koller M, et al. Oral biofilm analysis of palatal expanders by fluorescence in situ hybridization and confocal laser scanning microscopy[J]. J Vis Exp, 2011(56): 2967.
[20]	Hsu PP, Sabatini DM. Cancer cell metabolism: warburg and beyond[J]. Cell, 2008, 134(5): 703-707.
[21]	Sienski G, Dönertas D, Brennecke J. Transcriptional silencing of transposons by piwi and maelstrom and its impact on chromatin state and gene expression[J]. Cell, 2012, 151(5): 964-980.
[22]	Ocasio AB, Cotter PA. CDI/CDS system-encoding ge-nes of Burkholderia thailandensis are located in a mobile genetic element that defines a new class of transposon[J]. PLoS Genet, 2019, 15(1): e1007883.
[23]	Juhas M. Horizontal gene transfer in human pathogens[J]. Crit Rev Microbiol, 2015, 41(1): 101-108.
[24]	Davies EV, Winstanley C, Fothergill JL, et al. The role of temperate bacteriophages in bacterial infection[J]. FEMS Microbiol Lett, 2016, 363(5): fnw015.
[25]	Renard A, Diene SM, Courtier-Martinez L, et al. 12/111phiA prophage domestication is associated with autoaggregation and increased ability to produce biofilm in Streptococcus agalactiae [J]. Microorganisms, 2021, 9(6): 1112.
[26]	Han XL, Wang DY, Yang L, et al. Activation of polyamine catabolism promotes glutamine metabolism and creates a targetable vulnerability in lung cancer[J]. Proc Natl Acad Sci USA, 2024, 121(13): e2319429121.
[27]	Wang YX, Ledvina HE, Tower CA, et al. Discovery of a glutathione utilization pathway in Francisella that shows functional divergence between environmental and pathogenic species[J]. Cell Host Microbe, 2023, 31(8): 1359-1370.e7.
[28]	Wu HY, Yao Z, Li HK, et al. Improving dermal fibroblast-to-epidermis communications and aging wound repair through extracellular vesicle-mediated delivery of Gstm2 mRNA[J]. J Nanobiotechnol, 2024, 22: 307.

样品名称	浓度/（ng/μL）	总量/μg	OD_260/280	OD_260/230	RQN
野生型1	1 586.50	79.33	2.10	2.61	10.00
野生型2	1 691.20	59.19	2.11	2.59	9.90
野生型3	1 774.90	88.75	2.12	2.59	10.00
敲除株ΔgsT1	1 492.00	52.22	2.11	2.60	10.00
敲除株ΔgsT2	1 855.90	64.96	2.11	2.57	10.00
敲除株ΔgsT3	1 418.10	120.54	2.11	2.59	9.50

调节类型	FC>1.2	FC>1.5	FC>2
上调	99	22	8
下调	99	34	7