EspH utilizes phosphoinositide and Rab binding domains to interact with plasma membrane infection sites and Rab GTPases*

Enteropathogenic E. coli (EPEC) is a Gram-negative bacterial pathogen that causes persistent diarrhea. Upon attachment to the apical plasma membrane of the intestinal epithelium, the pathogen translocates virulence proteins called effectors into the infected cells. These effectors hijack numerous ho...

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Main Authors: Ipsita Nandi, Rachana Pattani Ramachandran, Deborah E. Shalev, Dina Schneidman-Duhovny, Raisa Shtuhin-Rahav, Naomi Melamed-Book, Efrat Zlotkin-Rivkin, Alexander Rouvinski, Ilan Rosenshine, Benjamin Aroeti
Format: Article
Language:English
Published: Taylor & Francis Group 2024-12-01
Series:Gut Microbes
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Online Access:https://www.tandfonline.com/doi/10.1080/19490976.2024.2400575
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author Ipsita Nandi
Rachana Pattani Ramachandran
Deborah E. Shalev
Dina Schneidman-Duhovny
Raisa Shtuhin-Rahav
Naomi Melamed-Book
Efrat Zlotkin-Rivkin
Alexander Rouvinski
Ilan Rosenshine
Benjamin Aroeti
author_facet Ipsita Nandi
Rachana Pattani Ramachandran
Deborah E. Shalev
Dina Schneidman-Duhovny
Raisa Shtuhin-Rahav
Naomi Melamed-Book
Efrat Zlotkin-Rivkin
Alexander Rouvinski
Ilan Rosenshine
Benjamin Aroeti
author_sort Ipsita Nandi
collection DOAJ
description Enteropathogenic E. coli (EPEC) is a Gram-negative bacterial pathogen that causes persistent diarrhea. Upon attachment to the apical plasma membrane of the intestinal epithelium, the pathogen translocates virulence proteins called effectors into the infected cells. These effectors hijack numerous host processes for the pathogen’s benefit. Therefore, studying the mechanisms underlying their action is crucial for a better understanding of the disease. We show that translocated EspH interacts with multiple host Rab GTPases. AlphaFold predictions and site-directed mutagenesis identified glutamic acid and lysine at positions 37 and 41 as Rab interacting residues in EspH. Mutating these sites abolished the ability of EspH to inhibit Akt and mTORC1 signaling, lysosomal exocytosis, and bacterial invasion. Knocking out the endogenous Rab8a gene expression highlighted the involvement of Rab8a in Akt/mTORC1 signaling and lysosomal exocytosis. A phosphoinositide binding domain with a critical tyrosine was identified in EspH. Mutating the tyrosine abolished the localization of EspH at infection sites and its capacity to interact with the Rabs. Our data suggest novel EspH-dependent mechanisms that elicit immune signaling and membrane trafficking during EPEC infection.
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spelling doaj-art-e905527152f04b9a97bcc2fed77a5ebc2025-08-20T02:16:55ZengTaylor & Francis GroupGut Microbes1949-09761949-09842024-12-0116110.1080/19490976.2024.2400575EspH utilizes phosphoinositide and Rab binding domains to interact with plasma membrane infection sites and Rab GTPases*Ipsita Nandi0Rachana Pattani Ramachandran1Deborah E. Shalev2Dina Schneidman-Duhovny3Raisa Shtuhin-Rahav4Naomi Melamed-Book5Efrat Zlotkin-Rivkin6Alexander Rouvinski7Ilan Rosenshine8Benjamin Aroeti9Department of Biological Chemistry, The Alexander Silberman Institute of Life Sciences, The Hebrew University of Jerusalem, The Edmond J. Safra Campus – Givat Ram, Jerusalem, IsraelDepartment of Biological Chemistry, The Alexander Silberman Institute of Life Sciences, The Hebrew University of Jerusalem, The Edmond J. Safra Campus – Givat Ram, Jerusalem, IsraelThe Wolfson Centre for Applied Structural Biology, The Hebrew University of Jerusalem, The Edmond J. Safra Campus – Givat Ram, Jerusalem, IsraelThe Rachel and Selim Benin School of Computer Science and Engineering, The Hebrew University of Jerusalem, Jerusalem, IsraelDepartment of Biological Chemistry, The Alexander Silberman Institute of Life Sciences, The Hebrew University of Jerusalem, The Edmond J. Safra Campus – Givat Ram, Jerusalem, IsraelBioimaging Unit, Alexander Silberman Institute of Life Sciences, The Hebrew University of Jerusalem, Jerusalem, IsraelDepartment of Biological Chemistry, The Alexander Silberman Institute of Life Sciences, The Hebrew University of Jerusalem, The Edmond J. Safra Campus – Givat Ram, Jerusalem, IsraelDepartment of Microbiology and Molecular Genetics, Institute for Medical Research Israel-Canada, Faculty of Medicine, The Hebrew University-Hadassah Medical School, of Jerusalem, Jerusalem, IsraelDepartment of Microbiology and Molecular Genetics, Institute for Medical Research Israel-Canada, Faculty of Medicine, The Hebrew University-Hadassah Medical School, of Jerusalem, Jerusalem, IsraelDepartment of Biological Chemistry, The Alexander Silberman Institute of Life Sciences, The Hebrew University of Jerusalem, The Edmond J. Safra Campus – Givat Ram, Jerusalem, IsraelEnteropathogenic E. coli (EPEC) is a Gram-negative bacterial pathogen that causes persistent diarrhea. Upon attachment to the apical plasma membrane of the intestinal epithelium, the pathogen translocates virulence proteins called effectors into the infected cells. These effectors hijack numerous host processes for the pathogen’s benefit. Therefore, studying the mechanisms underlying their action is crucial for a better understanding of the disease. We show that translocated EspH interacts with multiple host Rab GTPases. AlphaFold predictions and site-directed mutagenesis identified glutamic acid and lysine at positions 37 and 41 as Rab interacting residues in EspH. Mutating these sites abolished the ability of EspH to inhibit Akt and mTORC1 signaling, lysosomal exocytosis, and bacterial invasion. Knocking out the endogenous Rab8a gene expression highlighted the involvement of Rab8a in Akt/mTORC1 signaling and lysosomal exocytosis. A phosphoinositide binding domain with a critical tyrosine was identified in EspH. Mutating the tyrosine abolished the localization of EspH at infection sites and its capacity to interact with the Rabs. Our data suggest novel EspH-dependent mechanisms that elicit immune signaling and membrane trafficking during EPEC infection.https://www.tandfonline.com/doi/10.1080/19490976.2024.2400575Enteropathogenic e. colitype III secreted effectorsEspHRab GTPasesRho GTPasesphosphoinositides (PIs)
spellingShingle Ipsita Nandi
Rachana Pattani Ramachandran
Deborah E. Shalev
Dina Schneidman-Duhovny
Raisa Shtuhin-Rahav
Naomi Melamed-Book
Efrat Zlotkin-Rivkin
Alexander Rouvinski
Ilan Rosenshine
Benjamin Aroeti
EspH utilizes phosphoinositide and Rab binding domains to interact with plasma membrane infection sites and Rab GTPases*
Gut Microbes
Enteropathogenic e. coli
type III secreted effectors
EspH
Rab GTPases
Rho GTPases
phosphoinositides (PIs)
title EspH utilizes phosphoinositide and Rab binding domains to interact with plasma membrane infection sites and Rab GTPases*
title_full EspH utilizes phosphoinositide and Rab binding domains to interact with plasma membrane infection sites and Rab GTPases*
title_fullStr EspH utilizes phosphoinositide and Rab binding domains to interact with plasma membrane infection sites and Rab GTPases*
title_full_unstemmed EspH utilizes phosphoinositide and Rab binding domains to interact with plasma membrane infection sites and Rab GTPases*
title_short EspH utilizes phosphoinositide and Rab binding domains to interact with plasma membrane infection sites and Rab GTPases*
title_sort esph utilizes phosphoinositide and rab binding domains to interact with plasma membrane infection sites and rab gtpases
topic Enteropathogenic e. coli
type III secreted effectors
EspH
Rab GTPases
Rho GTPases
phosphoinositides (PIs)
url https://www.tandfonline.com/doi/10.1080/19490976.2024.2400575
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