A nonenzymatic effector disrupts <i>Bacteroides</i> cell wall homeostasis via OmpA targeting to mediate interbacterial competition

J Jing He Z Zhe Chen (Gladstone Institutes, San Francisco, CA, USA.) K Kun Jiang (Shanghai Key Laboratory of Molecular Catalysis and Innovative Materials, State Key Laboratory of Porous Materials for Separation and Conversion, iChEM (Collaborative Innovation Center of Chemistry for Energy Materials), Department of Chemistry) Y Yan Yang W Weixun Li (State Key Laboratory of Microbial Technology, Shandong University) X Xiaotong Wang (Department of Chemistry and Shanghai Key Laboratory of Molecular Catalysis and Innovative Materials, Fudan University, 220 Handan Road, Shanghai 200433, P. R. China) X Xiaoning Xu (State Key Laboratory of Microbial Technology, Shandong University) S Shuaining Zheng (State Key Laboratory of Microbial Technology, Shandong University) X Xuyao Jiao (State Key Laboratory of Microbial Technology, Shandong University) X Xudong Chen (Ministry of Education Key Laboratory of Protein Science, Tsinghua-Peking Center for Life Sciences, Beijing Advanced Innovation Center for Structural Biology, School of Life Sciences, Tsinghua University) L Liujie Huo (State Key Laboratory of Microbial Technology, Shandong University) B Bentley Lim (Department of Microbial Pathogenesis and Microbial Sciences Institute, Yale University School of Medicine) S Shuang-Jiang Liu (State Key Laboratory of Microbial Technology, Shandong University) X Xiang Gao

Abstract

The human gut microbiome is a dynamic ecosystem where bacteria engage in interspecies competition using molecular weapons such as the type VI secretion system (T6SS). Here, we characterize BteO–BtiO, a unique effector–immunity pair in Bacteroides fragilis that mediates antagonism via a nonenzymatic mechanism. Microscopy reveals that BteO exposure leads to cell elongation, membrane blebbing, and lysis in sensitive strains. Structural and biochemical analyses demonstrate that BteO disrupts cell wall homeostasis by binding to conserved C-terminal domains of OmpA-family proteins (OmpAs), which are critical for outer membrane integrity. The immunity protein BtiO neutralizes BteO by mimicking the OmpA-binding interface. We further show that bile salts enhance BteO-mediated killing in vitro and that BteO confers a competitive advantage in the mammalian gut. Remarkably, BteO exhibits broad-spectrum activity across Bacteroides species. These findings reveal a nonenzymatic strategy of bacterial antagonism and broaden our understanding of T6SS effector diversity within Bacteroides .

Article Details

Volume / Issue Vol. 122, Issue 41
Published October 14, 2025
ISSN 0027-8424
Publisher National Academy of Sciences

Authors (14)

J

Jing He

Z

Zhe Chen

Gladstone Institutes, San Francisco, CA, USA.

K

Kun Jiang

Shanghai Key Laboratory of Molecular Catalysis and Innovative Materials, State Key Laboratory of Porous Materials for Separation and Conversion, iChEM (Collaborative Innovation Center of Chemistry for Energy Materials), Department of Chemistry

Y

Yan Yang

W

Weixun Li

State Key Laboratory of Microbial Technology, Shandong University

X

Xiaotong Wang

Department of Chemistry and Shanghai Key Laboratory of Molecular Catalysis and Innovative Materials, Fudan University, 220 Handan Road, Shanghai 200433, P. R. China

X

Xiaoning Xu

State Key Laboratory of Microbial Technology, Shandong University

S

Shuaining Zheng

State Key Laboratory of Microbial Technology, Shandong University

X

Xuyao Jiao

State Key Laboratory of Microbial Technology, Shandong University

X

Xudong Chen

Ministry of Education Key Laboratory of Protein Science, Tsinghua-Peking Center for Life Sciences, Beijing Advanced Innovation Center for Structural Biology, School of Life Sciences, Tsinghua University

L

Liujie Huo

State Key Laboratory of Microbial Technology, Shandong University

B

Bentley Lim

Department of Microbial Pathogenesis and Microbial Sciences Institute, Yale University School of Medicine

S

Shuang-Jiang Liu

State Key Laboratory of Microbial Technology, Shandong University

X

Xiang Gao