title: Active Transport of Phosphorylated Carbohydrates Promotes Intestinal Colonization and Transmission of a Bacterial Pathogen.
abstract: Efficient acquisition of extracellular nutrients is essential for bacterial pathogenesis, however the identities and mechanisms for transport of many of these substrates remain unclear. Here, we investigate the predicted iron-binding transporter AfuABC and its role in bacterial pathogenesis in vivo. By crystallographic, biophysical and in vivo approaches, we show that AfuABC is in fact a cyclic hexose/heptose-phosphate transporter with high selectivity and specificity for a set of ubiquitous metabolites (glucose-6-phosphate, fructose-6-phosphate and sedoheptulose-7-phosphate). AfuABC is conserved across a wide range of bacterial genera, including the enteric pathogens EHEC O157:H7 and its murine-specific relative Citrobacter rodentium, where it lies adjacent to genes implicated in sugar sensing and acquisition. C. rodentium ΔafuA was significantly impaired in an in vivo murine competitive assay as well as its ability to transmit infection from an afflicted to a naïve murine host. Sugar-phosphates were present in normal and infected intestinal mucus and stool samples, indicating that these metabolites are available within the intestinal lumen for enteric bacteria to import during infection. Our study shows that AfuABC-dependent uptake of sugar-phosphates plays a critical role during enteric bacterial infection and uncovers previously unrecognized roles for these metabolites as important contributors to successful pathogenesis.
authors: Department of Biochemistry, University of Toronto, Toronto, Ontario, Canada.
authors: Brandon
authors: B
authors: Sit
authors: Department of Pediatrics and the Child and Family Research Institute, University of British Columbia, Vancouver, British Columbia, Canada.
authors: Shauna M
authors: SM
authors: Crowley
authors: Department of Pediatrics and the Child and Family Research Institute, University of British Columbia, Vancouver, British Columbia, Canada.
authors: Kirandeep
authors: K
authors: Bhullar
authors: Department of Biochemistry, University of Toronto, Toronto, Ontario, Canada.
authors: Christine Chieh-Lin
authors: CC
authors: Lai
authors: Department of Biochemistry, University of Toronto, Toronto, Ontario, Canada.
authors: Calvin
authors: C
authors: Tang
authors: Department of Biochemistry, University of Toronto, Toronto, Ontario, Canada.
authors: Yogesh
authors: Y
authors: Hooda
authors: Department of Biochemistry, University of Toronto, Toronto, Ontario, Canada.
authors: Charles
authors: C
authors: Calmettes
authors: Department of Microbiology and Infectious Diseases, University of Calgary, Calgary, Alberta, Canada; Department of Biochemistry and Molecular Biology, University of Calgary, Calgary, Alberta, Canada.
authors: Husain
authors: H
authors: Khambati
authors: Department of Pediatrics and the Child and Family Research Institute, University of British Columbia, Vancouver, British Columbia, Canada.
authors: Caixia
authors: C
authors: Ma
authors: Department of Molecular Genetics and Institute of Medical Science, University of Toronto, Ontario, Canada; Program in Cell Biology, Hospital for Sick Children, Toronto, Ontario, Canada; SickKids Inflammatory Bowel Disease Centre, Toronto, Ontario, Canada.
authors: John H
authors: JH
authors: Brumell
authors: Department of Microbiology and Infectious Diseases, University of Calgary, Calgary, Alberta, Canada; Department of Biochemistry and Molecular Biology, University of Calgary, Calgary, Alberta, Canada.
authors: Anthony B
authors: AB
authors: Schryvers
authors: Department of Pediatrics and the Child and Family Research Institute, University of British Columbia, Vancouver, British Columbia, Canada.
authors: Bruce A
authors: BA
authors: Vallance
authors: Department of Biochemistry, University of Toronto, Toronto, Ontario, Canada.
authors: Trevor F
authors: TF
authors: Moraes
mesh_terms: D000818:Animals; D001693:Biological Transport, Active / Q000502:physiology; D002151:Calorimetry; D050260:Carbohydrate Metabolism / Q000502:physiology*; D002853:Chromatography, Liquid; D044082:Citrobacter rodentium; D004756:Enterobacteriaceae Infections / Q000378:metabolism* / Q000635:transmission*; D007413:Intestinal Mucosa / Q000378:metabolism; D007422:Intestines / Q000382:microbiology*; D051379:Mice; D008810:Mice, Inbred C57BL; D008856:Microscopy, Fluorescence; D016297:Mutagenesis, Site-Directed; D010766:Phosphorylation; D010802:Phylogeny; D053719:Tandem Mass Spectrometry