Identification of NCAM that interacts with the PHE-CoV spike protein

Virol J. 2010 Sep 24:7:254. doi: 10.1186/1743-422X-7-254.

Abstract

Background: The spike proteins of coronaviruses associate with cellular molecules to mediate infection of their target cells. The characterization of cellular proteins required for virus infection is essential for understanding viral life cycles and may provide cellular targets for antiviral therapies.

Results: We identified Neural Cell Adhesion Molecule (NCAM) as a novel interacting partner of the PHE-CoV S protein. A T7 phage display cDNA library from N2a cells was constructed, and the library was screened with the soluble PHE-CoV S glycoproteins. We used a coimmunoprecipitation assay to show that only the NCAM was a binding partner of spike protein. We found that a soluble form of anti-NCAM antibody blocked association of the PHE-CoV with N2a cells. Furthermore, double-stranded siRNA targeted against NCAM inhibited PHE-CoV infection.

Conclusions: A novel interaction was identified between NCAM and spike protein and this association is critical during PHE-CoV infection.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cell Line
  • Coronavirus / pathogenicity*
  • Gene Silencing
  • Host-Pathogen Interactions*
  • Immunoprecipitation
  • Membrane Glycoproteins / metabolism*
  • Mice
  • Neural Cell Adhesion Molecules / metabolism*
  • Peptide Library
  • Podoviridae / genetics
  • Protein Binding
  • RNA, Small Interfering / genetics
  • RNA, Small Interfering / metabolism
  • Spike Glycoprotein, Coronavirus
  • Viral Envelope Proteins / metabolism*

Substances

  • Membrane Glycoproteins
  • Neural Cell Adhesion Molecules
  • Peptide Library
  • RNA, Small Interfering
  • Spike Glycoprotein, Coronavirus
  • Viral Envelope Proteins
  • spike glycoprotein, SARS-CoV
  • spike protein, mouse hepatitis virus