A genome-wide CRISPR screen identifies N-acetylglucosamine-1-phosphate transferase as a potential antiviral target for Ebola virus

Nat Commun. 2019 Jan 17;10(1):285. doi: 10.1038/s41467-018-08135-4.

Abstract

There are no approved therapies for Ebola virus infection. Here, to find potential therapeutic targets, we perform a screen for genes essential for Ebola virus (EBOV) infection. We identify GNPTAB, which encodes the α and β subunits of N-acetylglucosamine-1-phosphate transferase. We show that EBOV infection of a GNPTAB knockout cell line is impaired, and that this is reversed by reconstituting GNPTAB expression. Fibroblasts from patients with mucolipidosis II, a disorder associated with mutations in GNPTAB, are refractory to EBOV, whereas cells from their healthy parents support infection. Impaired infection correlates with loss of the expression of cathepsin B, known to be essential for EBOV entry. GNPTAB activity is dependent upon proteolytic cleavage by the SKI-1/S1P protease. Inhibiting this protease with the small-molecule PF-429242 blocks EBOV entry and infection. Disruption of GNPTAB function may represent a strategy for a host-targeted therapy for EBOV.

Publication types

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

MeSH terms

  • A549 Cells
  • Animals
  • Antiviral Agents / pharmacology*
  • Antiviral Agents / therapeutic use
  • Cathepsin B / metabolism
  • Chlorocebus aethiops
  • Clustered Regularly Interspaced Short Palindromic Repeats / genetics
  • Ebolavirus / pathogenicity*
  • Fibroblasts
  • Gene Knockout Techniques
  • Hemorrhagic Fever, Ebola / drug therapy*
  • Hemorrhagic Fever, Ebola / virology
  • Humans
  • Mucolipidoses / genetics
  • Mucolipidoses / pathology*
  • Proprotein Convertases / antagonists & inhibitors
  • Proprotein Convertases / metabolism
  • Protease Inhibitors / pharmacology
  • Pyrrolidines / pharmacology
  • Serine Endopeptidases / metabolism
  • Transferases (Other Substituted Phosphate Groups) / antagonists & inhibitors
  • Transferases (Other Substituted Phosphate Groups) / genetics*
  • Vero Cells
  • Virus Internalization / drug effects*
  • Whole Genome Sequencing

Substances

  • Antiviral Agents
  • PF-429242
  • Protease Inhibitors
  • Pyrrolidines
  • Transferases (Other Substituted Phosphate Groups)
  • GNPTAB protein, human
  • UDPacetylglucosamine-dolichyl-phosphate acetylglucosamine-1-phosphate transferase
  • Proprotein Convertases
  • Serine Endopeptidases
  • membrane-bound transcription factor peptidase, site 1
  • CTSB protein, human
  • Cathepsin B