Structure of a pentameric virion-associated fiber with a potential role in Orsay virus entry to host cells

PLoS Pathog. 2017 Feb 27;13(2):e1006231. doi: 10.1371/journal.ppat.1006231. eCollection 2017 Feb.

Abstract

Despite the wide use of Caenorhabditis elegans as a model organism, the first virus naturally infecting this organism was not discovered until six years ago. The Orsay virus and its related nematode viruses have a positive-sense RNA genome, encoding three proteins: CP, RdRP, and a novel δ protein that shares no homology with any other proteins. δ can be expressed either as a free δ or a CP-δ fusion protein by ribosomal frameshift, but the structure and function of both δ and CP-δ remain unknown. Using a combination of electron microscopy, X-ray crystallography, computational and biophysical analyses, here we show that the Orsay δ protein forms a ~420-Å long, pentameric fiber with an N-terminal α-helical bundle, a β-stranded filament in the middle, and a C-terminal head domain. The pentameric nature of the δ fiber has been independently confirmed by both mass spectrometry and analytical ultracentrifugation. Recombinant Orsay capsid containing CP-δ shows protruding long fibers with globular heads at the distal end. Mutant viruses with disrupted CP-δ fibers were generated by organism-based reverse genetics. These viruses were found to be either non-viable or with poor infectivity according to phenotypic and qRT-PCR analyses. Furthermore, addition of purified δ proteins to worm culture greatly reduced Orsay infectivity in a sequence-specific manner. Based on the structure resemblance between the Orsay CP-δ fiber and the fibers from reovirus and adenovirus, we propose that CP-δ functions as a cell attachment protein to mediate Orsay entry into worm intestine cells.

Publication types

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

MeSH terms

  • Animals
  • Caenorhabditis elegans / virology*
  • Capsid Proteins / chemistry
  • Capsid Proteins / ultrastructure*
  • Circular Dichroism
  • Crystallography, X-Ray
  • Mass Spectrometry
  • Microscopy, Electron, Transmission
  • Mutagenesis, Site-Directed
  • Organisms, Genetically Modified
  • RNA Virus Infections
  • RNA Viruses / physiology*
  • RNA Viruses / ultrastructure
  • Virion / chemistry
  • Virion / ultrastructure
  • Virus Internalization*

Substances

  • Capsid Proteins