title: Ebola virus entry requires the cholesterol transporter Niemann-Pick C1.
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.
authors.affiliation: Whitehead Institute for Biomedical Research, Nine Cambridge Center, Cambridge, Massachusetts 02142, USA.
authors.forename: Jan E
authors.initials: JE
authors.lastname: Carette
authors.forename: Matthijs
authors.initials: M
authors.lastname: Raaben
authors.forename: Anthony C
authors.initials: AC
authors.lastname: Wong
authors.forename: Andrew S
authors.initials: AS
authors.lastname: Herbert
authors.forename: Gregor
authors.initials: G
authors.lastname: Obernosterer
authors.forename: Nirupama
authors.initials: N
authors.lastname: Mulherkar
authors.forename: Ana I
authors.initials: AI
authors.lastname: Kuehne
authors.forename: Philip J
authors.initials: PJ
authors.lastname: Kranzusch
authors.forename: April M
authors.initials: AM
authors.lastname: Griffin
authors.forename: Gordon
authors.initials: G
authors.lastname: Ruthel
authors.forename: Paola
authors.initials: P
authors.lastname: Dal Cin
authors.forename: John M
authors.initials: JM
authors.lastname: Dye
authors.forename: Sean P
authors.initials: SP
authors.lastname: Whelan
authors.forename: Kartik
authors.initials: K
authors.lastname: Chandran
authors.forename: Thijn R
authors.initials: TR
authors.lastname: Brummelkamp
mesh_terms: D000818:Animals; D001692:Biological Transport; D002352:Carrier Proteins / Q000235:genetics / Q000378:metabolism*; D002460:Cell Line; D002784:Cholesterol / Q000378:metabolism*; D029043:Ebolavirus / Q000502:physiology*; D011992:Endosomes / Q000378:metabolism; D005347:Fibroblasts / Q000378:metabolism / Q000473:pathology / Q000821:virology; D015894:Genome, Human / Q000235:genetics; D006023:Glycoproteins / Q000378:metabolism; D006238:Haploidy; D019142:Hemorrhagic Fever, Ebola / Q000188:drug therapy / Q000378:metabolism; D054884:Host-Pathogen Interactions / Q000235:genetics; D006801:Humans; D047908:Intracellular Signaling Peptides and Proteins; D008247:Lysosomes / Q000378:metabolism; D008379:Marburg Virus Disease / Q000188:drug therapy / Q000378:metabolism; D029024:Marburgvirus / Q000502:physiology; D008561:Membrane Fusion / Q000235:genetics / Q000502:physiology; D008562:Membrane Glycoproteins / Q000172:deficiency / Q000235:genetics / Q000378:metabolism*; D046912:Multiprotein Complexes / Q000737:chemistry / Q000172:deficiency / Q000235:genetics / Q000378:metabolism; D009154:Mutation / Q000235:genetics; D000077296:Niemann-Pick C1 Protein; D009542:Niemann-Pick Diseases / Q000473:pathology / Q000821:virology; D011991:Receptors, Virus / Q000378:metabolism; D033921:Vesicular Transport Proteins; D014760:Viral Fusion Proteins / Q000378:metabolism; D053586:Virus Internalization
chemical_list: D002352:Carrier Proteins; D006023:Glycoproteins; D047908:Intracellular Signaling Peptides and Proteins; D008562:Membrane Glycoproteins; D046912:Multiprotein Complexes; C106881:NPC1 protein, human; C104585:NPC2 protein, human; D000077296:Niemann-Pick C1 Protein; D011991:Receptors, Virus; D033921:Vesicular Transport Proteins; D014760:Viral Fusion Proteins; D002784:Cholesterol