title: Intracellular Uropathogenic E. coli Exploits Host Rab35 for Iron Acquisition and Survival within Urinary Bladder Cells.
abstract: Recurrent urinary tract infections (UTIs) caused by uropathogenic E. coli (UPEC) are common and morbid infections with limited therapeutic options. Previous studies have demonstrated that persistent intracellular infection of bladder epithelial cells (BEC) by UPEC contributes to recurrent UTI in mouse models of infection. However, the mechanisms employed by UPEC to survive within BEC are incompletely understood. In this study we aimed to understand the role of host vesicular trafficking proteins in the intracellular survival of UPEC. Using a cell culture model of intracellular UPEC infection, we found that the small GTPase Rab35 facilitates UPEC survival in UPEC-containing vacuoles (UCV) within BEC. Rab35 plays a role in endosomal recycling of transferrin receptor (TfR), the key protein responsible for transferrin-mediated cellular iron uptake. UPEC enhance the expression of both Rab35 and TfR and recruit these proteins to the UCV, thereby supplying UPEC with the essential nutrient iron. Accordingly, Rab35 or TfR depleted cells showed significantly lower intracellular iron levels and reduced ability to support UPEC survival. In the absence of Rab35, UPEC are preferentially trafficked to degradative lysosomes and killed. Furthermore, in an in vivo murine model of persistent intracellular infection, Rab35 also colocalizes with intracellular UPEC. We propose a model in which UPEC subverts two different vesicular trafficking pathways (endosomal recycling and degradative lysosomal fusion) by modulating Rab35, thereby simultaneously enhancing iron acquisition and avoiding lysosomal degradation of the UCV within bladder epithelial cells. Our findings reveal a novel survival mechanism of intracellular UPEC and suggest a potential avenue for therapeutic intervention against recurrent UTI.
authors.affiliation: Program in Emerging Infectious Diseases, Duke-NUS Graduate Medical School, Singapore.
authors.forename: Neha
authors.initials: N
authors.lastname: Dikshit
authors.affiliation: Program in Emerging Infectious Diseases, Duke-NUS Graduate Medical School, Singapore.
authors.forename: Pradeep
authors.initials: P
authors.lastname: Bist
authors.affiliation: Infectious Diseases Group, Genome Institute of Singapore, Singapore; Faculty of Medicine and Institute for Life Sciences, University of Southampton, Southampton, United Kingdom.
authors.forename: Shannon N
authors.initials: SN
authors.lastname: Fenlon
authors.affiliation: Program in Emerging Infectious Diseases, Duke-NUS Graduate Medical School, Singapore.
authors.forename: Niyas Kudukkil
authors.initials: NK
authors.lastname: Pulloor
authors.affiliation: Department of Biochemistry, Yong Loo Lin School of Medicine, National University of Singapore, Singapore.
authors.forename: Christelle En Lin
authors.initials: CE
authors.lastname: Chua
authors.affiliation: Department of Microbiology and Immunology, College of Veterinary Medicine, Cornell University, Ithaca, New York, United States of America.
authors.forename: Marci A
authors.initials: MA
authors.lastname: Scidmore
authors.affiliation: Department of Microbiology and Immunology, Virginia Commonwealth University School of Medicine, Richmond, Virginia, United States of America.
authors.forename: Jason A
authors.initials: JA
authors.lastname: Carlyon
authors.affiliation: Department of Biochemistry, Yong Loo Lin School of Medicine, National University of Singapore, Singapore.
authors.forename: Bor Luen
authors.initials: BL
authors.lastname: Tang
authors.affiliation: Infectious Diseases Group, Genome Institute of Singapore, Singapore; Department of Medicine, Division of Infectious Diseases, Yong Loo Lin School of Medicine, National University of Singapore, Singapore.
authors.forename: Swaine L
authors.initials: SL
authors.lastname: Chen
authors.affiliation: Program in Emerging Infectious Diseases, Duke-NUS Graduate Medical School, Singapore.
authors.forename: Bindu
authors.initials: B
authors.lastname: Sukumaran
mesh_terms: D000111:Acetylcysteine; D000818:Animals; D002460:Cell Line; D004926:Escherichia coli / Q000378:metabolism; D004927:Escherichia coli Infections / Q000378:metabolism*; D005260:Female; D005455:Fluorescent Antibody Technique; D006790:Host-Parasite Interactions / Q000502:physiology*; D006801:Humans; D007501:Iron / Q000378:metabolism; D051379:Mice; D008810:Mice, Inbred C57BL; D018613:Microscopy, Confocal; D016133:Polymerase Chain Reaction; D021381:Protein Transport / Q000502:physiology; D014162:Transfection; D001743:Urinary Bladder / Q000382:microbiology; D014552:Urinary Tract Infections / Q000378:metabolism* / Q000382:microbiology; D056689:Uropathogenic Escherichia coli / Q000378:metabolism; D020691:rab GTP-Binding Proteins / Q000378:metabolism*
chemical_list: D007501:Iron; C511183:RAB35 protein, human; C543587:Rab35 protein, mouse; D020691:rab GTP-Binding Proteins; D000111:Acetylcysteine