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Related Experiment Videos

Shigella flexneri DegP facilitates IcsA surface expression and is required for efficient intercellular spread.

Georgiana E Purdy1, Mei Hong, Shelley M Payne

  • 1Institute for Cellular and Molecular Biology and Section of Molecular Genetics and Microbiology, The University of Texas at Austin, Austin, Texas 78712, USA.

Infection and Immunity
|October 16, 2002
PubMed
Summary
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A Shigella flexneri degP mutant showed reduced plaque formation due to impaired IcsA localization. DegP is crucial for bacterial virulence and survival under stress, but its protease activity is not essential for plaque formation.

Area of Science:

  • Microbiology
  • Bacterial Pathogenesis
  • Cell Biology

Background:

  • Shigella flexneri is an invasive bacterial pathogen responsible for bacillary dysentery.
  • Efficient intracellular spread and plaque formation are critical virulence factors for S. flexneri.
  • The protein IcsA (Invasive Plasmid antigen 1) is essential for bacterial motility and actin-based cell-to-cell spread.

Purpose of the Study:

  • To investigate the role of the DegP protease in the virulence of Shigella flexneri.
  • To determine the effect of degP mutation on bacterial invasion, plaque formation, and IcsA localization.
  • To assess the contribution of DegP's protease activity to S. flexneri virulence.

Main Methods:

  • Screening for insertion mutants with altered plaque formation.

Related Experiment Videos

  • Invasion assays using Henle and Caco-2 cell lines.
  • Analysis of IcsA localization using immunofluorescence microscopy.
  • Assessment of bacterial survival under stress conditions (elevated temperature, acidic pH).

    • Main Results:

    • A degP mutant (SM1100) exhibited reduced plaque size compared to wild-type S. flexneri.
    • The proportion of SM1100 bacteria with properly polarized IcsA was significantly reduced.
    • DegP's protease activity was not required for IcsA localization or plaque formation in Henle cells.
    • DegP was essential for efficient polar IcsA localization in E. coli expressing icsA.
    • The degP mutant showed compromised growth and survival at elevated temperatures and acidic conditions.

    Conclusions:

    • DegP plays a significant role in Shigella flexneri virulence, primarily by ensuring proper polar localization of the IcsA protein.
    • While DegP's protease activity is not essential for plaque formation, the protein itself is crucial for virulence and stress tolerance.
    • These findings highlight DegP as a potential target for therapeutic interventions against Shigella infections.