Role of lipid rafts in E-cadherin-- and HGF-R/Met--mediated entry of Listeria monocytogenes into host cells

The Journal of Cell Biology
Stéphanie SeveauPascale Cossart

Abstract

Listeria monocytogenes uptake by nonphagocytic cells is promoted by the bacterial invasion proteins internalin and InlB, which bind to their host receptors E-cadherin and hepatocyte growth factor receptor (HGF-R)/Met, respectively. Here, we present evidence that plasma membrane organization in lipid domains is critical for Listeria uptake. Cholesterol depletion by methyl-beta-cyclodextrin reversibly inhibited Listeria entry. Lipid raft markers, such as glycosylphosphatidylinositol-linked proteins, a myristoylated and palmitoylated peptide and the ganglioside GM1 were recruited at the bacterial entry site. We analyzed which molecular events require membrane cholesterol and found that the presence of E-cadherin in lipid domains was necessary for initial interaction with internalin to promote bacterial entry. In contrast, the initial interaction of InlB with HGF-R did not require membrane cholesterol, whereas downstream signaling leading to F-actin polymerization was cholesterol dependent. Our work, in addition to documenting for the first time the role of lipid rafts in Listeria entry, provides the first evidence that E-cadherin and HGF-R require lipid domain integrity for their full activity.

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Citations

Nov 25, 2006·Journal of the American Society for Mass Spectrometry·Koka JayasimhuluJosé-Luis Giner
Nov 2, 2004·Nature Cell Biology·Javier Pizarro-Cerdá, Pascale Cossart
May 20, 2006·Nature Reviews. Microbiology·Mélanie HamonPascale Cossart
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Jul 7, 2009·Annual Review of Cell and Developmental Biology·Javier Pizarro-Cerdá, Pascale Cossart
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Apr 12, 2005·Molecular Microbiology·Frank Lafont, F Gisou van der Goot
Oct 16, 2015·Developmental Cell·Alpha S YapRobert G Parton

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Methods Mentioned

BETA
myristoylation
fluorescence microscopy
coimmunoprecipitation
acylation
phase-contrast
immunoprecipitation

Software Mentioned

Metamorph
Adobe photoshop
Metamorph Imaging System

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