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The membrane localization domain is required for intracellular localization and autoregulation of YopE in Yersinia

Elin L Isaksson1, Margareta Aili, Anna Fahlgren

  • 1Department of Molecular Biology, Laboratory of Molecular Infectious Medicine, Umeå University, S-90187 Umeå, Sweden.

Infection and Immunity
|August 19, 2009
PubMed

Insights

The membrane localization domain (MLD) of YopE is crucial for targeting within host cells and bacterial virulence. Deleting the MLD impairs YopE

Area of Science:

  • Microbiology
  • Cell Biology
  • Molecular Biology

Background:

  • YopE is a key virulence factor of *Yersinia pseudotuberculosis*.
  • Proper localization of YopE in eukaryotic cells is essential for its function.
  • The membrane localization domain (MLD) of YopE (amino acids 54-75) has been implicated in YopE localization.

Purpose of the Study:

  • To investigate the role of the YopE MLD in *Yersinia pseudotuberculosis* virulence.
  • To determine the effect of MLD deletion on YopE secretion, translocation, localization, and function.

Main Methods:

  • Construction of an *in cis* MLD deletion mutant of YopE in *Y. pseudotuberculosis*.
  • Analysis of YopE secretion and translocation levels.
  • Assessment of YopE GTPase activating protein (GAP) activity towards Rac1 and RhoA.
  • Microscopic analysis of YopE intracellular localization in infected HeLa cells.
  • Evaluation of *Y. pseudotuberculosis* virulence using the MLD deletion mutant.

Main Results:

  • The YopE MLD deletion mutant secreted and translocated YopE at wild-type levels.
  • The mutant exhibited defective autoregulation of YopE expression and delayed cytotoxicity in HeLa cells.
  • GAP activity of the mutant YopE towards Rac1 and RhoA remained wild-type.
  • YopE lacking the MLD showed altered intracellular localization, dispersing throughout the cell, including the nucleus.
  • The YopE MLD deletion mutant was found to be avirulent in a mouse model.

Conclusions:

  • The MLD of YopE is essential for its proper intracellular targeting and virulence, independent of its GAP activity on Rac1 and RhoA.
  • YopE likely targets additional host proteins beyond Rac1 and RhoA for full virulence.
  • The MLD plays a critical role in mediating YopE's function during infection, suggesting it is a true virulence effector protein.

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