DNA Polymerase IV dinB Favors the Adaptive Fitness of mcr‐carrying Bacteria Through a Negative Feedback Regulatory Mechanism

Abstract The plasmid‐borne resistance gene mcr drastically undermines the effectiveness of colistin, posing a substantial threat to public health. Although several key plasmid elements that balance mcr‐1 persistence and bacterial growth are identified, the regulatory interactions between mcr‐1 and h...

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Main Authors: Haijie Zhang, Xia Xiao, Chenlong Wang, Yurong Zhao, Bo Chen, Xinyuan Ji, Lina Gu, Jie Wang, Zhiqiang Wang, Yuan Liu
Format: Article
Language:English
Published: Wiley 2025-03-01
Series:Advanced Science
Subjects:
Online Access:https://doi.org/10.1002/advs.202411994
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author Haijie Zhang
Xia Xiao
Chenlong Wang
Yurong Zhao
Bo Chen
Xinyuan Ji
Lina Gu
Jie Wang
Zhiqiang Wang
Yuan Liu
author_facet Haijie Zhang
Xia Xiao
Chenlong Wang
Yurong Zhao
Bo Chen
Xinyuan Ji
Lina Gu
Jie Wang
Zhiqiang Wang
Yuan Liu
author_sort Haijie Zhang
collection DOAJ
description Abstract The plasmid‐borne resistance gene mcr drastically undermines the effectiveness of colistin, posing a substantial threat to public health. Although several key plasmid elements that balance mcr‐1 persistence and bacterial growth are identified, the regulatory interactions between mcr‐1 and host bacteria remain poorly understood. Using a genome‐wide CRISPRi crRNA library, it is identified that DNA polymerase IV, dinB, is essential for controlling the fitness cost associated with mcr‐1 in Escherichia coli. The absence of dinB operon enhances mcr‐1‐mediated colistin resistance but simultaneously compromises bacterial growth and competitiveness. Meanwhile, dinB deficiency mitigates inflammatory response in RAW267.4 cells and enhances bacterial colonization in murine tissues. Further investigation reveals that mcr‐1 actively upregulates dinB expression, with the increased reactive oxygen species induced by mcr‐1 being crucial for this activation.   These findings suggest that dinB modulates mcr expression and bacterial fitness via a negative feedback regulatory mechanism. Leveraging this regulatory relationship, a Toxin‐Intein is engineered under the control of dinB promoter to selectively target and kill mcr‐positive E. coli both in vitro and in vivo. Overall, the work uncovers a novel adaptive mechanism underlying mcr persistence and provides a precise antimicrobial strategy to combat antibiotic‐resistant pathogens.
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spelling doaj-art-73ff19e3bf77472e9629adb5fb2f5d332025-08-20T01:49:41ZengWileyAdvanced Science2198-38442025-03-011212n/an/a10.1002/advs.202411994DNA Polymerase IV dinB Favors the Adaptive Fitness of mcr‐carrying Bacteria Through a Negative Feedback Regulatory MechanismHaijie Zhang0Xia Xiao1Chenlong Wang2Yurong Zhao3Bo Chen4Xinyuan Ji5Lina Gu6Jie Wang7Zhiqiang Wang8Yuan Liu9Jiangsu Co‐innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses College of Veterinary Medicine Yangzhou University Yangzhou 225009 ChinaJiangsu Co‐innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses College of Veterinary Medicine Yangzhou University Yangzhou 225009 ChinaJiangsu Co‐innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses College of Veterinary Medicine Yangzhou University Yangzhou 225009 ChinaJiangsu Co‐innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses College of Veterinary Medicine Yangzhou University Yangzhou 225009 ChinaJiangsu Co‐innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses College of Veterinary Medicine Yangzhou University Yangzhou 225009 ChinaJiangsu Co‐innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses College of Veterinary Medicine Yangzhou University Yangzhou 225009 ChinaJiangsu Co‐innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses College of Veterinary Medicine Yangzhou University Yangzhou 225009 ChinaJiangsu Co‐innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses College of Veterinary Medicine Yangzhou University Yangzhou 225009 ChinaJiangsu Co‐innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses College of Veterinary Medicine Yangzhou University Yangzhou 225009 ChinaJiangsu Co‐innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses College of Veterinary Medicine Yangzhou University Yangzhou 225009 ChinaAbstract The plasmid‐borne resistance gene mcr drastically undermines the effectiveness of colistin, posing a substantial threat to public health. Although several key plasmid elements that balance mcr‐1 persistence and bacterial growth are identified, the regulatory interactions between mcr‐1 and host bacteria remain poorly understood. Using a genome‐wide CRISPRi crRNA library, it is identified that DNA polymerase IV, dinB, is essential for controlling the fitness cost associated with mcr‐1 in Escherichia coli. The absence of dinB operon enhances mcr‐1‐mediated colistin resistance but simultaneously compromises bacterial growth and competitiveness. Meanwhile, dinB deficiency mitigates inflammatory response in RAW267.4 cells and enhances bacterial colonization in murine tissues. Further investigation reveals that mcr‐1 actively upregulates dinB expression, with the increased reactive oxygen species induced by mcr‐1 being crucial for this activation.   These findings suggest that dinB modulates mcr expression and bacterial fitness via a negative feedback regulatory mechanism. Leveraging this regulatory relationship, a Toxin‐Intein is engineered under the control of dinB promoter to selectively target and kill mcr‐positive E. coli both in vitro and in vivo. Overall, the work uncovers a novel adaptive mechanism underlying mcr persistence and provides a precise antimicrobial strategy to combat antibiotic‐resistant pathogens.https://doi.org/10.1002/advs.202411994bacteriacolistin resistancedinBfitness costmcr expression
spellingShingle Haijie Zhang
Xia Xiao
Chenlong Wang
Yurong Zhao
Bo Chen
Xinyuan Ji
Lina Gu
Jie Wang
Zhiqiang Wang
Yuan Liu
DNA Polymerase IV dinB Favors the Adaptive Fitness of mcr‐carrying Bacteria Through a Negative Feedback Regulatory Mechanism
Advanced Science
bacteria
colistin resistance
dinB
fitness cost
mcr expression
title DNA Polymerase IV dinB Favors the Adaptive Fitness of mcr‐carrying Bacteria Through a Negative Feedback Regulatory Mechanism
title_full DNA Polymerase IV dinB Favors the Adaptive Fitness of mcr‐carrying Bacteria Through a Negative Feedback Regulatory Mechanism
title_fullStr DNA Polymerase IV dinB Favors the Adaptive Fitness of mcr‐carrying Bacteria Through a Negative Feedback Regulatory Mechanism
title_full_unstemmed DNA Polymerase IV dinB Favors the Adaptive Fitness of mcr‐carrying Bacteria Through a Negative Feedback Regulatory Mechanism
title_short DNA Polymerase IV dinB Favors the Adaptive Fitness of mcr‐carrying Bacteria Through a Negative Feedback Regulatory Mechanism
title_sort dna polymerase iv dinb favors the adaptive fitness of mcr carrying bacteria through a negative feedback regulatory mechanism
topic bacteria
colistin resistance
dinB
fitness cost
mcr expression
url https://doi.org/10.1002/advs.202411994
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