Spontaneous mutation at amino acid 544 of the Ebola virus glycoprotein potentiates virus entry and selection in tissue culture

John B. Ruedas, Jason T. Ladner, Chelsea R. Ettinger, Suryaram Gummuluru, Gustavo Palacios, John H. Connor

Research output: Contribution to journalArticlepeer-review

17 Scopus citations


Ebolaviruses have a surface glycoprotein (GP1,2) that is required for virus attachment and entry into cells. Mutations affecting GP1,2 functions can alter virus growth properties. We generated a recombinant vesicular stomatitis virus encoding Ebola virus Makona variant GP1,2 (rVSV-MAK-GP) and observed emergence of a T544I mutation in the Makona GP1,2 gene during tissue culture passage in certain cell lines. The T544I mutation emerged within two passages when VSV-MAK-GP was grown on Vero E6, Vero, and BS-C-1 cells but not when it was passaged on Huh7 and HepG2 cells. The mutation led to a marked increase in virus growth kinetics and conferred a robust growth advantage over wild-type rVSV-MAK-GP on Vero E6 cells. Analysis of complete viral genomes collected from patients in western Africa indicated that this mutation was not found in Ebola virus clinical samples. However, we observed the emergence of T544I during serial passage of various Ebola Makona isolates on Vero E6 cells. Three independent isolates showed emergence of T544I from undetectable levels in nonpassaged virus or virus passaged once to frequencies of greater than 60% within a single passage, consistent with it being a tissue culture adaptation. Intriguingly, T544I is not found in any Sudan, Bundibugyo, or Tai Forest ebolavirus sequences. Furthermore, T544I did not emerge when we serially passaged recombinant VSV encoding GP1,2 from these ebolaviruses. This report provides experimental evidence that the spontaneous mutation T544I is a tissue culture adaptation in certain cell lines and that it may be unique for the species Zaire ebolavirus.

Original languageEnglish (US)
Article numbere00392-17
JournalJournal of Virology
Issue number15
StatePublished - Aug 1 2017
Externally publishedYes


  • Adaptation
  • Bundibugyo ebolavirus
  • Ebola virus
  • Glycoproteins
  • Membrane fusion
  • Mutation
  • Sudan ebolavirus
  • Tai Forest ebolavirus
  • Tissue culture
  • Tissue culture mutation
  • Virus entry

ASJC Scopus subject areas

  • Microbiology
  • Immunology
  • Insect Science
  • Virology


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