The Pseudomonas aeruginosa type III secretion system delivers effector proteins directly into target cells, allowing the bacterium to modulate host cell functions. ExoU is the most cytotoxic of the known effector proteins and has been associated with more severe infections in humans. Previous studies have shown that ExoU is a patatin-like A2 phospholipase requiring the cellular host factors phosphatidylinositol 4,5-bisphosphate (PI(4,5)P2) and ubiquitin for its activation in vitro. We demonstrated that PI(4,5)P2 also induces oligomerization of ExoU and that this PI(4,5)P2-mediated oligomerization does not require ubiquitin. Single amino acid substitutions in the C-terminal membrane localization domain of ExoU reduced both its activity as well as its ability to form higher-order complexes in transfected cells and in vitro. Combining inactive truncated ExoU proteins partially restored phospholipase activity and cytotoxicity, indicating that ExoU oligomerization may have functional significance. Our results indicate that PI(4,5)P2 induces oligomerization of ExoU, which may be a mechanism by which this co-activator enhances the phospholipase activity of ExoU.

Last modified

• 01/18/2019

Creator

• Angelica Zhang

DOI

• https://doi.org/10.21985/N21B4S

Subject

• Driskill Graduate Training Program in Life Sciences

Keyword

• Pseudomonas aeruginosa
• ExoU
• oligomerization

Date created

• 2017-01-01

Resource type

• Dissertation

Rights statement

• In Copyright