Elucidation of the Ebola virus VP24 cellular interactome and disruption of virus biology through targeted inhibition of host-cell protein function

J Proteome Res. 2014 Nov 7;13(11):5120-35. doi: 10.1021/pr500556d. Epub 2014 Oct 23.

Abstract

Viral pathogenesis in the infected cell is a balance between antiviral responses and subversion of host-cell processes. Many viral proteins specifically interact with host-cell proteins to promote virus biology. Understanding these interactions can lead to knowledge gains about infection and provide potential targets for antiviral therapy. One such virus is Ebola, which has profound consequences for human health and causes viral hemorrhagic fever where case fatality rates can approach 90%. The Ebola virus VP24 protein plays a critical role in the evasion of the host immune response and is likely to interact with multiple cellular proteins. To map these interactions and better understand the potential functions of VP24, label-free quantitative proteomics was used to identify cellular proteins that had a high probability of forming the VP24 cellular interactome. Several known interactions were confirmed, thus placing confidence in the technique, but new interactions were also discovered including one with ATP1A1, which is involved in osmoregulation and cell signaling. Disrupting the activity of ATP1A1 in Ebola-virus-infected cells with a small molecule inhibitor resulted in a decrease in progeny virus, thus illustrating how quantitative proteomics can be used to identify potential therapeutic targets.

Keywords: Ebola virus; VP24 protein; antiviral; inhibitor; interactome; label free proteomics; proteomics; virus.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cell Line / drug effects
  • Cell Line / virology
  • Ebolavirus / drug effects
  • Ebolavirus / pathogenicity*
  • Green Fluorescent Proteins / genetics
  • Green Fluorescent Proteins / metabolism
  • HEK293 Cells / drug effects
  • HEK293 Cells / virology
  • Host-Pathogen Interactions
  • Humans
  • Mass Spectrometry / methods
  • Ouabain / pharmacology
  • Protein Interaction Mapping / methods*
  • Proteomics / methods
  • Reproducibility of Results
  • Sodium-Potassium-Exchanging ATPase / antagonists & inhibitors
  • Sodium-Potassium-Exchanging ATPase / metabolism*
  • Viral Proteins / genetics
  • Viral Proteins / metabolism*

Substances

  • VP24 protein, Ebola virus
  • Viral Proteins
  • enhanced green fluorescent protein
  • Green Fluorescent Proteins
  • Ouabain
  • ATP1A1 protein, human
  • Sodium-Potassium-Exchanging ATPase