Disruption of Ebola NP0VP35 Inclusion Body-like Structures reduce Viral Infection

J Mol Biol. 2023 Oct 15;435(20):168241. doi: 10.1016/j.jmb.2023.168241. Epub 2023 Aug 19.

Abstract

Viral inclusion bodies (IBs) are potential sites of viral replication and assembly. How viral IBs form remains poorly defined. Here we describe a combined biophysical and cellular approach to identify the components necessary for IB formation during Ebola virus (EBOV) infection. We find that the eNP0VP35 complex containing Ebola nucleoprotein (eNP) and viral protein 35 (eVP35), the functional equivalents of nucleoprotein (N) and phosphoprotein (P) in non-segmented negative strand viruses (NNSVs), phase separates to form inclusion bodies. Phase separation of eNP0VP35 is reversible and modulated by ionic strength. The multivalency of eVP35, and not eNP, is also critical for phase separation. Furthermore, overexpression of an eVP35 peptide disrupts eNP0VP35 complex formation, leading to reduced frequency of IB formation and limited viral infection. Together, our results show that upon EBOV infection, the eNP0VP35 complex forms the minimum unit to drive IB formation and viral replication.

Keywords: Ebola; biomolecular condensate; inclusion body; phase separation; viral replication organelle.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Ebolavirus* / metabolism
  • Ebolavirus* / physiology
  • Hemorrhagic Fever, Ebola* / virology
  • Humans
  • Inclusion Bodies* / virology
  • Nucleoproteins* / metabolism
  • Viral Regulatory and Accessory Proteins / metabolism
  • Virus Replication*

Substances

  • Nucleoproteins
  • Viral Regulatory and Accessory Proteins
  • nucleoprotein VP35, Ebola virus