Disruptions in outer membrane-peptidoglycan interactions enhance bile salt resistance in O-antigen-producing E. coli

mBio

卷:16

Research Article

Jilong Qin^1,2,3  Yaoqin Hong^1,2,4  Waldemar Vollmer^3,5  Renato Morona⁶  Makrina Totsika^1,2 

[1] School of Biological Sciences, Department of Molecular & Biomedical Sciences, Research Centre for Infectious Diseases, University of Adelaide, Adelaide, South Australia, Australia

[2] Centre for Immunology and Infection Control, School of Biomedical Sciences, Queensland University of Technology, Brisbane, Queensland, Australia

[3] Institute for Molecular Bioscience, The University of Queensland, Brisbane, Queensland, Australia

[4] Biomedical Sciences and Molecular Biology, College of Medicine and Dentistry, James Cook University, Townsville City, Queensland, Australia

[5] Max Planck Queensland Centre, Queensland University of Technology, Brisbane, Queensland, Australia

[6] Centre for Bacterial Cell Biology, Biosciences Institute, Newcastle University, Newcastle upon Tyne, United Kingdom

关键词: polysaccharides;    cell envelope;    O antigen;    peptidoglycan;    Lpp;    OmpA;   

DOI  :  10.1128/mbio.02184-25

 received in 2025-07-16, accepted in 2025-08-01,  发布年份 2025

来源: American Society for Microbiology

【 摘 要 】

ABSTRACTBile salts (BS) are antimicrobials that disrupt bacterial cell membranes and induce oxidative stress. The gut bacterium Escherichia coli is naturally resistant to BS, including the model strain K12 MG1655 that produces a lipopolysaccharide (LPS) without O-antigen (OAg) on the cell surface. Paradoxically, we have previously shown that restoring a wild-type like LPS with attached OAg (MG1655-S) sensitizes E. coli K12 to exogenous BS. In this study, we investigate this phenomenon. We show that mutations causing truncation of the LPS core oligosaccharide render MG1655-S strains even more susceptible to BS compared to MG1655. These mutants phenocopy a K-12 MG1655-S ΔwaaL mutant, which is defective in OAg ligase, primarily due to periplasmic accumulation of the unligated lipid-linked UndPP-OAg. Through the characterization of BS-resistant suppressor mutants of MG1655-S ΔwaaL, we identify key genetic disruptions involved in resistance. Notably, we observed the highest BS resistance in strains with a weaker connection between the outer membrane (OM) and peptidoglycan (PG), including strains lacking the major OM-anchored, PG-binding proteins OmpA or Lpp. Expressing versions of OmpA and Lpp that lack PG-binding capacity also enhanced the BS resistance. Our data suggest that BS-induced stress in OAg-producing E. coli is due to the spatial constraints between OM and PG and that mutations disrupting OM–PG interactions alleviate this stress, thereby enhancing BS resistance. These findings provide new insights into a major challenge E. coli faces in the gut environment, where it needs to produce OAg for stable colonization and immune evasion while resisting the antimicrobial activity of BS.IMPORTANCEEnteric bacteria residing in the human gut must withstand host-derived antimicrobial bile salts, but resistance mechanisms are not fully elucidated. In this study, we investigate bile salt resistance mechanisms in O-antigen (OAg)-producing Escherichia coli K-12. We show that the accumulation of carrier lipid-linked OAg in the periplasm of strains with truncated lipopolysaccharide (LPS) core oligosaccharide or defects in OAg ligase can sensitize E. coli more to bile salt, unless the physical links between outer membrane and peptidoglycan are disrupted, highlighting that bile salt-induced stress is attributed to spatial constraints between the outer membrane and peptidoglycan layer. Our work uncovers a previously unappreciated envelope stress response mechanism in E. coli, where reducing outer membrane–peptidoglycan connectivity mitigates bile salt-induced damage arising from OAg production. These findings reshape our understanding of how physical architecture and biosynthetic intermediates intersect to influence bacterial survival in hostile environments.

【 授权许可】

CC BY   
Copyright © 2025 Qin et al.

【 图 表 】

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