Journal article
Targeting of microvillus protein Eps8 by the NleH effector kinases from enteropathogenic E. coli
GL Pollock, AM Grishin, C Giogha, J Gan, CV Oates, PJ McMillan, I Gaeta, MJ Tyska, JS Pearson, NE Scott, M Cygler, EL Hartland
Proceedings of the National Academy of Sciences of the United States of America | Published : 2022
Abstract
Attaching and effacing (AE) lesion formation on enterocytes by enteropathogenic Escherichia coli (EPEC) requires the EPEC type III secretion system (T3SS). Two T3SS effectors injected into the host cell during infection are the atypical kinases, NleH1 and NleH2. However, the host targets of NleH1 and NleH2 kinase activity during infection have not been reported. Here phosphoproteomics identified Ser775 in the microvillus protein Eps8 as a bona fide target of NleH1 and NleH2 phosphorylation. Both kinases interacted with Eps8 through previously unrecognized, noncanonical "proline-rich"motifs, PxxDY, that bound the Src Homology 3 (SH3) domain of Eps8. Structural analysis of the Eps8 SH3 domain ..
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Awarded by University of Saskatchewan
Funding Acknowledgements
This work was supported by National Health and Medical Research Council of Australia project grants awarded to E.L.H. (APP1175976). N.E.S. was supported by an Overseas (Biomedical) Fellowship (APP1037373) and is currently the recipient of an Australian Research Council Future Fellowship (FT200100270). G.L.P. was the recipient of an Australian Postgraduate Award. J.G. was supported by a China Scholarship Council-University of Melbourne PhD scholarship. I.G. was supported by the Vanderbilt Cellular, Biochemical and Molecular Sciences training grant 5T32GM 008554-25. M.J.T. was supported by National Institutes of Health grants R01-DK111949 and R01-DK095811. The crystallography diffraction dataset described in this paper was collected from beamline Canadian Macromolecular Crystallography Facility-Bending Magnet at the Canadian Light Source, a national research facility of the University of Saskatchewan, which is supported by the Canada Foundation for Innovation, the Natural Sciences and Engineering Research Council, the National Research Council, the Canadian Institutes of Health Research, the Government of Saskatchewan, and the University of Saskatchewan. The data integration script XDSGO was provided by Dr. Denis Spasyuk. We gratefully acknowledge the Protein Characterization and Crystallization Facility, College of Medicine, University of Saskatchewan, funded by the Canadian Foundation for Innovation and facility manager Dr. Michal Boniecki, for help with access to specialized instrumentation. Super resolution microscopy imaging was performed at the Biological Optical Microscopy Platform at the University of Melbourne. The authors acknowledge Monash Micro Imaging, Monash University, for the provision of instrumentation, training, and technical support.