A Hyperstabilizing Mutation in the Base of the Ebola Virus Glycoprotein Acts at Multiple Steps To Abrogate Viral Entry

J. Maximilian Fels, Jennifer S. Spence, Robert H. Bortz, Zachary A. Bornholdt, Kartik Chandran

Research output: Contribution to journalArticle

Abstract

Ebola virus (EBOV) causes highly lethal disease outbreaks against which no FDA-approved countermeasures are available. Although many host factors exploited by EBOV for cell entry have been identified, including host cell surface phosphatidylserine receptors, endosomal cysteine proteases, and the lysosomal cholesterol trafficking protein NPC1, key questions remain. Specifically, late entry steps culminating in viral membrane fusion remain enigmatic. Here, we investigated a set of glycoprotein (GP) mutants previously hypothesized to be entry defective and identified one mutation, R64A, that abolished infection with no apparent impact on GP expression, folding, or viral incorporation. R64A profoundly thermostabilized EBOV GP and rendered it highly resistant to proteolysis in vitro Forward-genetics and cell entry studies strongly suggested that R64A's effects on GP thermostability and proteolysis arrest viral entry at least at two distinct steps: the first upstream of NPC1 binding and the second at a late entry step downstream of fusion activation. Concordantly, toremifene, a small-molecule entry inhibitor previously shown to bind and destabilize GP, may selectively enhance the infectivity of viral particles bearing GP(R64A) at subinhibitory concentrations. R64A provides a valuable tool to further define the interplay between GP stability, proteolysis, and viral membrane fusion; to explore the rational design of stability-modulating antivirals; and to spur the development of next-generation Ebola virus vaccines with improved stability.IMPORTANCE Ebola virus is a medically relevant virus responsible for outbreaks of severe disease in western and central Africa, with mortality rates reaching as high as 90%. Despite considerable effort, there are currently no FDA-approved therapeutics or targeted interventions available, highlighting the need of development in this area. Host-cell invasion represents an attractive target for antivirals, and several drug candidates have been identified; however, our limited understanding of the complex viral entry process challenges the development of such entry-targeting drugs. Here, we report on a glycoprotein mutation that abrogates viral entry and provides insights into the final steps of this process. In addition, the hyperstabilized phenotype of this mutant makes it useful as a tool in the discovery and design of stability-modulating antivirals and next-generation vaccines against Ebola virus.

Original languageEnglish (US)
JournalmBio
Volume10
Issue number4
DOIs
StatePublished - Jul 9 2019

Fingerprint

Ebolavirus
Glycoproteins
Mutation
Virus Internalization
Ebola Vaccines
Proteolysis
Antiviral Agents
Disease Outbreaks
Toremifene
Central Africa
Western Africa
Cysteine Proteases
Cell Surface Receptors
Protein Transport
Drug Delivery Systems
Virion
Cholesterol
Viruses
Phenotype
Mortality

Keywords

  • cell entry
  • Ebola
  • ebolavirus
  • filovirus
  • fusion activation
  • glycoprotein
  • thermostability
  • viral entry
  • viral membrane fusion

ASJC Scopus subject areas

  • Microbiology
  • Virology

Cite this

A Hyperstabilizing Mutation in the Base of the Ebola Virus Glycoprotein Acts at Multiple Steps To Abrogate Viral Entry. / Fels, J. Maximilian; Spence, Jennifer S.; Bortz, Robert H.; Bornholdt, Zachary A.; Chandran, Kartik.

In: mBio, Vol. 10, No. 4, 09.07.2019.

Research output: Contribution to journalArticle

Fels, J. Maximilian ; Spence, Jennifer S. ; Bortz, Robert H. ; Bornholdt, Zachary A. ; Chandran, Kartik. / A Hyperstabilizing Mutation in the Base of the Ebola Virus Glycoprotein Acts at Multiple Steps To Abrogate Viral Entry. In: mBio. 2019 ; Vol. 10, No. 4.
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