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Mutations in the clpS Gene of Riemerella anatipestifer Affect Stress Response and Bacterial Virulence

Liu Yufu1,2,3, Liu Jiayi4, Liao Ruixin4

  • 1School of Life Sciences, Zhaoqing University, Zhaoqing, 526061, Guangdong, China. 363995606@qq.com.

Current Microbiology
|November 24, 2025
PubMed

Insights

The ClpS protein is crucial for Riemerella anatipestifer survival under stress and for its virulence. Deleting the clpS gene impairs bacterial stress response, adhesion, invasion, and pathogenicity in ducklings.

Area of Science:

  • Microbiology
  • Molecular Biology
  • Pathogenesis

Background:

  • The Clp protease system, particularly the ClpS subunit, plays a vital role in microbial post-transcriptional regulation, affecting antibiotic resistance, biofilm formation, stress response, and virulence.
  • The specific function of ClpS in Riemerella anatipestifer, a significant veterinary pathogen, has not been previously elucidated.

Purpose of the Study:

  • To investigate the role of the clpS gene in Riemerella anatipestifer.
  • To understand the impact of clpS deletion on bacterial survival, stress response, virulence, and gene expression.

Main Methods:

  • Construction of an in-frame deletion mutant (ΔclpS) of Riemerella anatipestifer using homologous recombination.
  • Assessment of bacterial growth rate, morphology, and survival under various stress conditions (heat, oxidative, acidic, iron deficiency).
  • Evaluation of adhesion and invasion capabilities using RAW264.7 cells and pathogenicity in a duckling model.
  • RNA sequencing (RNA-seq) analysis to identify differentially expressed genes in the ΔclpS mutant.

Main Results:

  • The ΔclpS mutant showed no significant changes in growth rate or morphology but exhibited significantly reduced survival under heat, oxidative, acidic, and iron-deficient conditions.
  • Riemerella anatipestifer activates clpS transcription under heat and oxidative stress.
  • The ΔclpS mutant displayed impaired adhesion and invasion of host cells and reduced pathogenicity in ducklings.
  • RNA-seq analysis revealed differential expression of 77 genes in the ΔclpS mutant, primarily affecting metabolic pathways.

Conclusions:

  • The clpS gene is essential for Riemerella anatipestifer's stress response and virulence.
  • ClpS contributes to bacterial survival under various environmental stresses and plays a role in host-pathogen interactions.
  • These findings provide insights into the molecular mechanisms underlying Riemerella anatipestifer pathogenesis.

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