GSBS Dissertations and Theses

Approval Date

4-25-2014

Document Type

Doctoral Dissertation

Academic Program

MD/PhD

Department

Molecular Genetics and Microbiology

First Thesis Advisor

John M. Leong, MD, PhD

Keywords

Actin Cytoskeleton, Actins, Calpain, Cytoskeletal Proteins, Enterohemorrhagic Escherichia coli, Escherichia coli Infections, Escherichia coli Proteins, Microvilli

Subjects

Dissertations, UMMS; Actin Cytoskeleton; Actins; Calpain; Cytoskeletal Proteins; Enterohemorrhagic Escherichia coli; Escherichia coli Infections; Escherichia coli Proteins; Microvilli

Abstract

Enterohemorrhagic E. coli (EHEC) is a major cause of food borne diarrheal illness worldwide. While disease symptoms are usually self-resolving and limited to severe gastroenteritis with bloody diarrhea, EHEC infection can lead to a life threatening complication known as Hemolytic Uremic Syndrome (HUS), which strikes children disproportionately and is the leading cause of kidney failure in children. Upon infection of gut epithelia, EHEC produces characteristic lesions called actin pedestals. These striking formations involve dramatic rearrangement of host cytoskeletal proteins. EHEC hijacks mammalian signaling pathways to cause destruction of microvilli and rebuilds the actin cytoskeleton underneath sites of bacterial attachment. Here, we present a brief study on a host factor, Calpain, involved in microvilli effacement, and an in depth investigation on a bacterial factor, EspFU, required for actin pedestal formation in intestinal cell models. Calpain is activated by both EHEC and the related pathogen, enteropathogenic E. coli (EPEC), during infection and facilitates microvilli disassembly by cleavage of a key membrane-cytoskeleton anchoring substrate, Ezrin. Actin pedestal formation is facilitated by the injection of two bacterial effectors, Tir and EspFU, into host cells, which work in concert to manipulate the host actin nucleators N-WASP and Arp2/3. EspFU hijacks key host signaling proteins N-WASP and IRTKS by mimetic displacement and has evolved to outcompete mammalian host ligands. Multiple repeats of key functional domains of EspFU are essential for actin pedestal activity through proper localization and competition against the an abundant host factor Eps8 for binding to IRTKS.

Comments

Copyright by (Cindy) YuShuan Lai 2014 All Rights Reserved

DOI

10.13028/M2HW2H

Rights and Permissions

Copyright is held by the author, with all rights reserved.

 
 

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