Ebola virus entry requires the cholesterol transporter Niemann-Pick C1

Nature. 2011 Aug 24;477(7364):340-3. doi: 10.1038/nature10348.

Abstract

Infections by the Ebola and Marburg filoviruses cause a rapidly fatal haemorrhagic fever in humans for which no approved antivirals are available. Filovirus entry is mediated by the viral spike glycoprotein (GP), which attaches viral particles to the cell surface, delivers them to endosomes and catalyses fusion between viral and endosomal membranes. Additional host factors in the endosomal compartment are probably required for viral membrane fusion; however, despite considerable efforts, these critical host factors have defied molecular identification. Here we describe a genome-wide haploid genetic screen in human cells to identify host factors required for Ebola virus entry. Our screen uncovered 67 mutations disrupting all six members of the homotypic fusion and vacuole protein-sorting (HOPS) multisubunit tethering complex, which is involved in the fusion of endosomes to lysosomes, and 39 independent mutations that disrupt the endo/lysosomal cholesterol transporter protein Niemann-Pick C1 (NPC1). Cells defective for the HOPS complex or NPC1 function, including primary fibroblasts derived from human Niemann-Pick type C1 disease patients, are resistant to infection by Ebola virus and Marburg virus, but remain fully susceptible to a suite of unrelated viruses. We show that membrane fusion mediated by filovirus glycoproteins and viral escape from the vesicular compartment require the NPC1 protein, independent of its known function in cholesterol transport. Our findings uncover unique features of the entry pathway used by filoviruses and indicate potential antiviral strategies to combat these deadly agents.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Animals
  • Biological Transport
  • Carrier Proteins / genetics
  • Carrier Proteins / metabolism*
  • Cell Line
  • Cholesterol / metabolism*
  • Ebolavirus / physiology*
  • Endosomes / metabolism
  • Fibroblasts / metabolism
  • Fibroblasts / pathology
  • Fibroblasts / virology
  • Genome, Human / genetics
  • Glycoproteins / metabolism
  • Haploidy
  • Hemorrhagic Fever, Ebola / drug therapy
  • Hemorrhagic Fever, Ebola / metabolism
  • Host-Pathogen Interactions / genetics
  • Humans
  • Lysosomes / metabolism
  • Marburg Virus Disease / drug therapy
  • Marburg Virus Disease / metabolism
  • Marburgvirus / physiology
  • Membrane Fusion / genetics
  • Membrane Fusion / physiology
  • Membrane Glycoproteins / deficiency
  • Membrane Glycoproteins / genetics
  • Membrane Glycoproteins / metabolism*
  • Multiprotein Complexes / chemistry
  • Multiprotein Complexes / deficiency
  • Multiprotein Complexes / genetics
  • Multiprotein Complexes / metabolism
  • Mutation / genetics
  • Niemann-Pick Diseases / pathology
  • Niemann-Pick Diseases / virology
  • Receptors, Virus / metabolism
  • Viral Fusion Proteins / metabolism
  • Virus Internalization*

Substances

  • Carrier Proteins
  • Glycoproteins
  • Membrane Glycoproteins
  • Multiprotein Complexes
  • NPC1 protein, human
  • NPC2 protein, human
  • Receptors, Virus
  • Viral Fusion Proteins
  • Cholesterol