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

Overview

Journal iScience

Publisher Cell Press

Date 2019 May 21

PMID 31108395

Citations 30

Authors

Toshiyuki Yamaji

Hisatoshi Hanamatsu

Tsuyoshi Sekizuka

Makoto Kuroda

Norimasa Iwasaki

Makoto Ohnishi

Jun-Ichi Furukawa

Kinnosuke Yahiro

Kentaro Hanada

Affiliations

Soon will be listed here.

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.

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