Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2024 Jan-Dec;16(1):2400575.
doi: 10.1080/19490976.2024.2400575. Epub 2024 Sep 23.

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

Ipsita Nandi  1   2 , Rachana Pattani Ramachandran  1   2 , Deborah E Shalev  3   4 , Dina Schneidman-Duhovny  5 , Raisa Shtuhin-Rahav  1   2 , Naomi Melamed-Book  6 , Efrat Zlotkin-Rivkin  1   2 , Alexander Rouvinski  7 , Ilan Rosenshine  7 , Benjamin Aroeti  1   2
Affiliations

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

Ipsita Nandi et al. Gut Microbes. 2024 Jan-Dec.

Abstract

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.

Keywords: Enteropathogenic e. coli; EspH; Rab GTPases; Rho GTPases; bacterial invasion; host-pathogen interactions; lysosomal exocytosis; phosphoinositide binding domain (PBD) PI3K/Akt/mTORC1 signaling; phosphoinositides (PIs); type III secreted effectors.

PubMed Disclaimer

Conflict of interest statement

No potential conflict of interest was reported by the author(s).

Figures

Figure 1.
Figure 1.
Rab8a, Rab10, Rab3a and Rab12 co-precipitate with translocated EspHwt.
Figure 2.
Figure 2.
Translocated EspHwt interacts with active Rab8a. Analysis by co-precipitation.
Figure 3.
Figure 3.
AlphaFold predicted structures for binding interfaces of EspH-Rabs.
Figure 4.
Figure 4.
The predicted Rab binding residues in EspH are critical for EspH-Rab interactions and are highly susceptible to mutagenesis.
Figure 5.
Figure 5.
The Rab binding residues in EspH are critical for exerting Rab-related functions.
Figure 6.
Figure 6.
Interactions of EspH with Rab8a are essential for eliciting the AkT/mTORC1 signaling, lysosomal exocytosis, and the interactions with Rab3a.
Figure 7.
Figure 7.
The effects of mutations in EspH used to study the interactions with Rab GTPases on host cell cytotoxicity and filopodia formation.
Figure 8.
Figure 8.
The EspH PBD is critical for EspH localization at infection sites, PI clustering, and interactions with Rab8a.
See this image and copyright information in PMC

Similar articles

See all similar articles

References

    1. Nataro JP, Kaper JB.. Diarrheagenic Escherichia coli. Clin Microbiol Rev. 1998;11(1):142–19. doi:10.1128/CMR.11.1.142. - DOI - PMC - PubMed
    1. Ochoa TJ, Contreras CA.. Enteropathogenic Escherichia coli infection in children. Curr Opin Infect Dis. 2011;24(5):478–483. doi:10.1097/QCO.0b013e32834a8b8b. - DOI - PMC - PubMed
    1. Kaur P, Dudeja PK. Pathophysiology of enteropathogenic Escherichia coli-induced diarrhea. Newborn. 2023;2(1):102–113. doi:10.5005/jp-journals-11002-0056. - DOI - PMC - PubMed
    1. Silberger DJ, Zindl CL, Weaver CT. Citrobacter rodentium: a model enteropathogen for understanding the interplay of innate and adaptive components of type 3 immunity. Mucosal Immunol. 2017;10(5):1108–1117. doi:10.1038/mi.2017.47. - DOI - PMC - PubMed
    1. Slater SL, Sagfors AM, Pollard DJ, Ruano-Gallego D, Frankel G. The type III secretion system of pathogenic Escherichia coli. Curr Top Microbiol Immunol. 2018;416:51–72. - PubMed

MeSH terms

Substances

Grants and funding

RRS and IN are the recipients of Dr. Willem Been Legacy Fellowship. This research was supported by the Israel Science Foundation [grant No. 1671/19].

LinkOut - more resources