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A CRISPR Screen Using Subtilase Cytotoxin Identifies SLC39A9 as a Glycan-Regulating Factor.

Authors
  • Yamaji, Toshiyuki1
  • Hanamatsu, Hisatoshi2
  • Sekizuka, Tsuyoshi3
  • Kuroda, Makoto3
  • Iwasaki, Norimasa4
  • Ohnishi, Makoto5
  • Furukawa, Jun-Ichi6
  • Yahiro, Kinnosuke7
  • Hanada, Kentaro8
  • 1 Department of Biochemistry and Cell Biology, National Institute of Infectious Diseases, Shinjuku-ku, Tokyo 162-8640, Japan. Electronic address: [email protected] , (Japan)
  • 2 Department of Advanced Clinical Glycobiology, Faculty of Medicine and Graduate School of Medicine, Hokkaido University, Sapporo 001-0021, Japan; Department of Gastroenterology and Hepatology, Graduate School of Medicine, Hokkaido University, Sapporo 060-8638, Japan. , (Japan)
  • 3 Pathogen Genomics Center, National Institute of Infectious Diseases, Shinjuku-ku, Tokyo 162-8640, Japan. , (Japan)
  • 4 Department of Advanced Clinical Glycobiology, Faculty of Medicine and Graduate School of Medicine, Hokkaido University, Sapporo 001-0021, Japan; Department of Orthopaedic Surgery, Graduate School of Medicine, Hokkaido University, Sapporo 060-8638, Japan. , (Japan)
  • 5 Department of Bacteriology I, National Institute of Infectious Diseases, Shinjuku-ku, Tokyo 162-8640, Japan. , (Japan)
  • 6 Department of Advanced Clinical Glycobiology, Faculty of Medicine and Graduate School of Medicine, Hokkaido University, Sapporo 001-0021, Japan. , (Japan)
  • 7 Department of Molecular Infectiology, Graduate School of Medicine, Chiba University, Chiba 260-8670, Japan. Electronic address: [email protected] , (Japan)
  • 8 Department of Biochemistry and Cell Biology, National Institute of Infectious Diseases, Shinjuku-ku, Tokyo 162-8640, Japan. , (Japan)
Type
Published Article
Journal
iScience
Publication Date
May 31, 2019
Volume
15
Pages
407–420
Identifiers
DOI: 10.1016/j.isci.2019.05.005
PMID: 31108395
Source
Medline
Keywords
Language
English
License
Unknown

Abstract

Subtilase cytotoxin (SubAB) is a virulence factor produced by locus of enterocyte effacement-negative Shiga-toxigenic Escherichia coli strains. The toxin recognizes sialoglycans for entry and cleaves an endoplasmic reticulum chaperon, binding immunoglobulin protein, to cause cell death. However, no systematic screening has yet been performed to identify critical host factors. Here, we performed a genome-wide CRISPR/Cas9 knockout screen for SubAB-induced cell death and identified various sialoglycan-related and membrane-trafficking genes. Analysis of glycan-deficient cells demonstrated that not only N-glycans but also O-glycans serve as SubAB receptors. In addition, SLC39A9, which is a predicted zinc transporter, as well as KDELRs and JTB, were required for SubAB to induce maximal cell death. Disruption of the SLC39A9 gene markedly reduced both complex-type N-glycans and core 1 O-glycans, and the O-glycan reduction was attributed to the reduction of core 1 synthase (C1GalT1). These results provide insights into the post-transcriptional regulation of glycosyltransferases by SLC39A9, as well as sialoglycan species as SubAB receptors. Copyright © 2019 The Author(s). Published by Elsevier Inc. All rights reserved.

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