Sodium butyrate inhibits the expression of virulence factors in Vibrio cholerae by targeting ToxT protein

ABSTRACT Cholera, a diarrheal disease caused by the gram-negative bacterium Vibrio cholerae, remains a global health threat in developing countries due to its high transmissibility and increased antibiotic resistance. There is a pressing need for alternative strategies, with an emphasis on anti-viru...

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Main Authors: Sushmita Kundu, Suman Das, Priyanka Maitra, Prolay Halder, Hemanta Koley, Asish K. Mukhopadhyay, Shin-ichi Miyoshi, Shanta Dutta, Nabendu Sekhar Chatterjee, Sushmita Bhattacharya
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Language:English
Published: American Society for Microbiology 2025-05-01
Series:mSphere
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Online Access:https://journals.asm.org/doi/10.1128/msphere.00824-24
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author Sushmita Kundu
Suman Das
Priyanka Maitra
Prolay Halder
Hemanta Koley
Asish K. Mukhopadhyay
Shin-ichi Miyoshi
Shanta Dutta
Nabendu Sekhar Chatterjee
Sushmita Bhattacharya
author_facet Sushmita Kundu
Suman Das
Priyanka Maitra
Prolay Halder
Hemanta Koley
Asish K. Mukhopadhyay
Shin-ichi Miyoshi
Shanta Dutta
Nabendu Sekhar Chatterjee
Sushmita Bhattacharya
author_sort Sushmita Kundu
collection DOAJ
description ABSTRACT Cholera, a diarrheal disease caused by the gram-negative bacterium Vibrio cholerae, remains a global health threat in developing countries due to its high transmissibility and increased antibiotic resistance. There is a pressing need for alternative strategies, with an emphasis on anti-virulent approaches to alter the outcome of bacterial infections, given the increase in antimicrobial-resistant strains. V. cholerae causes cholera by secreting virulence factors in the intestinal epithelial cells. These virulence factors facilitate bacterial colonization and cholera toxin production during infection. Here, we demonstrate that sodium butyrate (SB), a small molecule, had no effect on bacterial viability but was effective in suppressing the virulence attributes of V. cholerae. The production of cholera toxin (CT) was significantly reduced in a standard V. cholerae El Tor strain and two clinical isolates when grown in the presence of SB. Analysis of mRNA and protein levels further revealed that SB reduced the expression of the ToxT-dependent virulence genes like tcpA and ctxAB. DNA-protein interaction assays, conducted at cellular (ChIP) and in vitro conditions (EMSA), indicated that SB weakens the binding between ToxT and its downstream promoter DNA, likely by blocking DNA binding. Furthermore, the anti-virulence efficacy of SB was confirmed in animal models. These findings suggest that SB could be developed as an anti-virulence agent against V. cholerae, serving as a potential alternative to conventional antibiotics or as an adjunctive therapy to combat cholera.IMPORTANCEThe world has been facing an upsurge in cholera cases since 2021, a similar trend continuing into 2022, with over 29 countries reporting cholera outbreaks (World Health Organization, 16 December 2022, Disease Outbreak News, Cholera—global situation). Treatment of cholera involves oral rehydration therapy coupled with antibiotics to reduce the duration of the illness. However, in recent years, indiscriminate use of antibiotics has contributed to the emergence of antibiotic-resistant strains. In this study, we have addressed the problem of antibiotic resistance by targeting virulence factors. Screening various compounds using in silico methods led to the identification of a small molecule, SB, that inhibits the virulence cascade in V. cholerae. We demonstrated that (i) SB intervened in ToxT protein-DNA binding and subsequently affected the expression of ToxT-regulated virulence genes (ctxAB and tcpA) and (ii) SB is a potential therapeutic candidate for the development of a novel antimicrobial agent.
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spelling doaj-art-d6af8c1c73f942e6b1dd85f9cc390d922025-08-20T03:14:38ZengAmerican Society for MicrobiologymSphere2379-50422025-05-0110510.1128/msphere.00824-24Sodium butyrate inhibits the expression of virulence factors in Vibrio cholerae by targeting ToxT proteinSushmita Kundu0Suman Das1Priyanka Maitra2Prolay Halder3Hemanta Koley4Asish K. Mukhopadhyay5Shin-ichi Miyoshi6Shanta Dutta7Nabendu Sekhar Chatterjee8Sushmita Bhattacharya9Division of Biochemistry, ICMR-National Institute for Research in Bacterial Infections (Formerly ICMR-National Institute of Cholera and Enteric Diseases), Kolkata, IndiaDivision of Biochemistry, ICMR-National Institute for Research in Bacterial Infections (Formerly ICMR-National Institute of Cholera and Enteric Diseases), Kolkata, IndiaDivision of Biochemistry, ICMR-National Institute for Research in Bacterial Infections (Formerly ICMR-National Institute of Cholera and Enteric Diseases), Kolkata, IndiaDivision of Bacteriology, ICMR-National Institute for Research in Bacterial Infections (Formerly ICMR-National Institute of Cholera and Enteric Diseases), Kolkata, IndiaDivision of Bacteriology, ICMR-National Institute for Research in Bacterial Infections (Formerly ICMR-National Institute of Cholera and Enteric Diseases), Kolkata, IndiaDivision of Bacteriology, ICMR-National Institute for Research in Bacterial Infections (Formerly ICMR-National Institute of Cholera and Enteric Diseases), Kolkata, IndiaDivision of Pharmaceutical Sciences, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University, Okayama, JapanDivision of Bacteriology, ICMR-National Institute for Research in Bacterial Infections (Formerly ICMR-National Institute of Cholera and Enteric Diseases), Kolkata, IndiaDivision of Biochemistry, ICMR-National Institute for Research in Bacterial Infections (Formerly ICMR-National Institute of Cholera and Enteric Diseases), Kolkata, IndiaDivision of Biochemistry, ICMR-National Institute for Research in Bacterial Infections (Formerly ICMR-National Institute of Cholera and Enteric Diseases), Kolkata, IndiaABSTRACT Cholera, a diarrheal disease caused by the gram-negative bacterium Vibrio cholerae, remains a global health threat in developing countries due to its high transmissibility and increased antibiotic resistance. There is a pressing need for alternative strategies, with an emphasis on anti-virulent approaches to alter the outcome of bacterial infections, given the increase in antimicrobial-resistant strains. V. cholerae causes cholera by secreting virulence factors in the intestinal epithelial cells. These virulence factors facilitate bacterial colonization and cholera toxin production during infection. Here, we demonstrate that sodium butyrate (SB), a small molecule, had no effect on bacterial viability but was effective in suppressing the virulence attributes of V. cholerae. The production of cholera toxin (CT) was significantly reduced in a standard V. cholerae El Tor strain and two clinical isolates when grown in the presence of SB. Analysis of mRNA and protein levels further revealed that SB reduced the expression of the ToxT-dependent virulence genes like tcpA and ctxAB. DNA-protein interaction assays, conducted at cellular (ChIP) and in vitro conditions (EMSA), indicated that SB weakens the binding between ToxT and its downstream promoter DNA, likely by blocking DNA binding. Furthermore, the anti-virulence efficacy of SB was confirmed in animal models. These findings suggest that SB could be developed as an anti-virulence agent against V. cholerae, serving as a potential alternative to conventional antibiotics or as an adjunctive therapy to combat cholera.IMPORTANCEThe world has been facing an upsurge in cholera cases since 2021, a similar trend continuing into 2022, with over 29 countries reporting cholera outbreaks (World Health Organization, 16 December 2022, Disease Outbreak News, Cholera—global situation). Treatment of cholera involves oral rehydration therapy coupled with antibiotics to reduce the duration of the illness. However, in recent years, indiscriminate use of antibiotics has contributed to the emergence of antibiotic-resistant strains. In this study, we have addressed the problem of antibiotic resistance by targeting virulence factors. Screening various compounds using in silico methods led to the identification of a small molecule, SB, that inhibits the virulence cascade in V. cholerae. We demonstrated that (i) SB intervened in ToxT protein-DNA binding and subsequently affected the expression of ToxT-regulated virulence genes (ctxAB and tcpA) and (ii) SB is a potential therapeutic candidate for the development of a novel antimicrobial agent.https://journals.asm.org/doi/10.1128/msphere.00824-24sodium butyrate (SB)inhibitorpathogenesisVibrio choleraectxABantimicrobial resistance
spellingShingle Sushmita Kundu
Suman Das
Priyanka Maitra
Prolay Halder
Hemanta Koley
Asish K. Mukhopadhyay
Shin-ichi Miyoshi
Shanta Dutta
Nabendu Sekhar Chatterjee
Sushmita Bhattacharya
Sodium butyrate inhibits the expression of virulence factors in Vibrio cholerae by targeting ToxT protein
mSphere
sodium butyrate (SB)
inhibitor
pathogenesis
Vibrio cholerae
ctxAB
antimicrobial resistance
title Sodium butyrate inhibits the expression of virulence factors in Vibrio cholerae by targeting ToxT protein
title_full Sodium butyrate inhibits the expression of virulence factors in Vibrio cholerae by targeting ToxT protein
title_fullStr Sodium butyrate inhibits the expression of virulence factors in Vibrio cholerae by targeting ToxT protein
title_full_unstemmed Sodium butyrate inhibits the expression of virulence factors in Vibrio cholerae by targeting ToxT protein
title_short Sodium butyrate inhibits the expression of virulence factors in Vibrio cholerae by targeting ToxT protein
title_sort sodium butyrate inhibits the expression of virulence factors in vibrio cholerae by targeting toxt protein
topic sodium butyrate (SB)
inhibitor
pathogenesis
Vibrio cholerae
ctxAB
antimicrobial resistance
url https://journals.asm.org/doi/10.1128/msphere.00824-24
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