The two-component system CrdRS mediates sub-inhibitory antibiotic-induced biofilm formation in Helicobacter pylori via bidirectional regulation of transporters PlpA and GlnP
- 1Key Laboratory for Experimental Teratology of the Ministry of Education and Department of Microbiology, School of Basic Medical Sciences, Cheeloo College of Medicine, Shandong University, Jinan, Shandong, China
- 2Department of Pathology, School of Basic Medical Sciences, Cheeloo College of Medicine, Shandong University, Jinan, Shandong, China
Abstract
Biofilm formation by Helicobacter pylori is a major driver of antibiotic tolerance and treatment failure, yet the signaling pathways that trigger biofilm development under sub-inhibitory antibiotic pressure remain poorly understood. Here, we show that sub-MIC levels of metronidazole, amoxicillin, and ciprofloxacin potently induce dense H. pylori biofilms. Through transcriptomic and genetic analyses, we identify the two-component system CrdRS as a central signaling hub that orchestrates this response via the bidirectional regulation of two ATP-binding cassette (ABC) transporters. Phosphorylated CrdR binds to AC-rich promoter motifs to directly activate plpA, which encodes a substrate-binding protein that drives exopolysaccharide secretion and matrix assembly. Concomitantly, CrdR represses glnP, which encodes an inner-membrane permease, thereby relieving transcriptional inhibition of the L-asparaginase gene ansB. This derepression triggers aberrant reactive oxygen species (ROS) accumulation, which promotes oxidative stress-dependent biofilm maturation. Through phenotypic analysis of crdRS deletion mutants, phosphorylation-defective point mutants (CrdRD53A and CrdSH173A), and exogenous hydrogen peroxide (H₂O₂) induction, we demonstrate that CrdRS is required for mediating antibiotic-induced stress responses in a manner genetically separable from ROS sensing. Collectively, our findings establish a dual-mechanism model in which CrdRS orchestrates antibiotic-induced biofilm formation by simultaneously controlling matrix production and intracellular ROS generation. Notably, glnP expression was significantly lower in clinical multidrug-resistant isolates than in drug-sensitive ones, whereas plpA showed the opposite trend. These findings provide a mechanistic foundation for developing CrdRS-targeted strategies to combat biofilm-associated H. pylori infections.
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BibTeX
@article{zhang2026twocomponent,
title = {{The two-component system CrdRS mediates sub-inhibitory antibiotic-induced biofilm formation in Helicobacter pylori via bidirectional regulation of transporters PlpA and GlnP}},
author = {Wenxin Zhang and Lu Zhang and Yanlin Sun and Mingzhong Zhao and Shujing Xu and Han Yu and Wenjing Wang and Jinmeng Liu and Wenyue Ma and Xiaoyu Wang and Ziyan Zhang and Yundong Sun},
journal = {Antimicrobial Agents and Chemotherapy},
year = {2026},
month = oct,
doi = {10.1128/aac.00882-26},
url = {https://doi.org/10.1128/aac.00882-26},
note = {CC BY 4.0},
}
RIS
TY - JOUR
TI - The two-component system CrdRS mediates sub-inhibitory antibiotic-induced biofilm formation in Helicobacter pylori via bidirectional regulation of transporters PlpA and GlnP
AU - Wenxin Zhang
AU - Lu Zhang
AU - Yanlin Sun
AU - Mingzhong Zhao
AU - Shujing Xu
AU - Han Yu
AU - Wenjing Wang
AU - Jinmeng Liu
AU - Wenyue Ma
AU - Xiaoyu Wang
AU - Ziyan Zhang
AU - Yundong Sun
PY - 2026
DA - 2026/10/05
JO - Antimicrobial Agents and Chemotherapy
DO - 10.1128/aac.00882-26
UR - https://doi.org/10.1128/aac.00882-26
AB - Biofilm formation by Helicobacter pylori is a major driver of antibiotic tolerance and treatment failure, yet the signaling pathways that trigger biofilm development under sub-inhibitory antibiotic pressure remain poorly understood. Here, we show that sub-MIC levels of metronidazole, amoxicillin, and ciprofloxacin potently induce dense H. pylori biofilms. Through transcriptomic and genetic analyses, we identify the two-component system CrdRS as a central signaling hub that orchestrates this response via the bidirectional regulation of two ATP-binding cassette (ABC) transporters. Phosphorylated CrdR binds to AC-rich promoter motifs to directly activate plpA, which encodes a substrate-binding protein that drives exopolysaccharide secretion and matrix assembly. Concomitantly, CrdR represses glnP, which encodes an inner-membrane permease, thereby relieving transcriptional inhibition of the L-asparaginase gene ansB. This derepression triggers aberrant reactive oxygen species (ROS) accumulation, which promotes oxidative stress-dependent biofilm maturation. Through phenotypic analysis of crdRS deletion mutants, phosphorylation-defective point mutants (CrdRD53A and CrdSH173A), and exogenous hydrogen peroxide (H₂O₂) induction, we demonstrate that CrdRS is required for mediating antibiotic-induced stress responses in a manner genetically separable from ROS sensing. Collectively, our findings establish a dual-mechanism model in which CrdRS orchestrates antibiotic-induced biofilm formation by simultaneously controlling matrix production and intracellular ROS generation. Notably, glnP expression was significantly lower in clinical multidrug-resistant isolates than in drug-sensitive ones, whereas plpA showed the opposite trend. These findings provide a mechanistic foundation for developing CrdRS-targeted strategies to combat biofilm-associated H. pylori infections.
SN - 0066-4804
SN - 1098-6596
ER -