Abstract
Bacterial toxins require localization to specific intracellular compartments following injection into host cells. In this study, we examined the membrane targeting of a broad family of bacterial proteins, the patatin-like phospholipases. The best characterized member of this family is ExoU, an effector of the Pseudomonas aeruginosa type III secretion system. Upon injection into host cells, ExoU localizes to the plasma membrane, where it uses its phospholipase A2 activity to lyse infected cells. The targeting mechanism of ExoU is poorly characterized, but it was recently found to bind to the phospholipid phosphatidylinositol 4,5-bisphosphate (PI(4,5)P2), a marker for the plasma membrane of eukaryotic cells. We confirmed that the membrane localization domain (MLD) of ExoU had a direct affinity for PI(4,5)P2, and we determined that this binding was required for ExoU localization. Previously uncharacterized ExoU homologs from Pseudomonas fluorescens and Photorhabdus asymbiotica also localized to the plasma membrane and required PI(4,5)P2 for this localization. A conserved arginine within the MLD was critical for interaction of each protein with PI(4,5)P2 and for localization. Furthermore, we determined the crystal structure of the full-length P. fluorescens ExoU and found that it was similar to that of P. aeruginosa ExoU. Each MLD contains a fourhelical bundle, with the conserved arginine exposed at its cap to allow for interaction with the negatively charged PI(4,5)P2. Overall, these findings provide a structural explanation for the targeting of patatin-like phospholipases to the plasma membrane and define the MLD of ExoU as a member of a new class of PI(4,5)P2 binding domains.
Original language | English (US) |
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Pages (from-to) | 2919-2937 |
Number of pages | 19 |
Journal | Journal of Biological Chemistry |
Volume | 290 |
Issue number | 5 |
DOIs | |
State | Published - Jan 30 2015 |
ASJC Scopus subject areas
- Molecular Biology
- Biochemistry
- Cell Biology
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Crystal structure of type III effector protein ExoU (exoU)
Tyson, G. H. (Contributor), Halavaty, A. S. (Contributor), Kim, H. (Contributor), Geissler, B. (Contributor), Agard, M. (Contributor), Satchell, K. J. (Contributor), Cho, W. (Contributor), Anderson, W. F. (Contributor) & Hauser, A. R. (Contributor), Protein Data Bank (PDB), Dec 17 2014
DOI: 10.2210/pdb4QMK/pdb, https://www.wwpdb.org/pdb?id=pdb_00004qmk
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