title: Newcastle disease virus HN protein alters the conformation of the F protein at cell surfaces.
abstract: Conformational changes in the Newcastle disease virus (NDV) fusion (F) protein during activation of fusion and the role of HN protein in these changes were characterized with a polyclonal antibody. This antibody was raised against a peptide with the sequence of the amino-terminal half of the F protein HR1 domain. This antibody immunoprecipitated both F(0) and F(1) forms of the fusion protein from infected and transfected cell extracts solubilized with detergent, and precipitation was unaffected by expression of the HN protein. In marked contrast, this antibody detected significant conformational differences in the F protein at cell surfaces, differences that depended upon HN protein expression. The antibody minimally detected the F protein, either cleaved or uncleaved, in the absence of HN protein expression. However, when coexpressed with HN protein, an uncleaved mutant F protein bound the anti-HR1 antibody, and this binding depended upon the coexpression of specifically the NDV HN protein. When the cleaved wild-type F protein was coexpressed with HN protein, the F protein bound anti-HR1 antibody poorly although significantly more than F protein expressed alone. Anti-HR1 antibody inhibited the fusion of R18 (octadecyl rhodamine B chloride)-labeled red blood cells to syncytia expressing HN and wild-type F proteins. This inhibition showed that fusion-competent F proteins present on surfaces of syncytia were capable of binding anti-HR1. Furthermore, only antibody which was added prior to red blood cell binding could inhibit fusion. These results suggest that the conformation of uncleaved cell surface F protein is affected by HN protein expression. Furthermore, the cleaved F protein, when coexpressed with HN protein and in a prefusion conformation, can bind anti-HR1 antibody, and the anti-HR1-accessible conformation exists prior to HN protein attachment to receptors on red blood cells.
authors.affiliation: Department of Molecular Genetics and Microbiology/Program in Immunology and Virology, University of Massachusetts Medical School, 55 Lake Avenue North, Worcester, MA 01655.
authors.forename: Lori W
authors.initials: LW
authors.lastname: McGinnes
authors.forename: Kathryn
authors.initials: K
authors.lastname: Gravel
authors.forename: Trudy G
authors.initials: TG
authors.lastname: Morrison
mesh_terms: D000595:Amino Acid Sequence; D005455:Fluorescent Antibody Technique; D015707:HN Protein / Q000502:physiology*; D008561:Membrane Fusion; D008969:Molecular Sequence Data; D009522:Newcastle disease virus / Q000737:chemistry*; D011233:Precipitin Tests; D011487:Protein Conformation; D014760:Viral Fusion Proteins / Q000737:chemistry*
chemical_list: D015707:HN Protein; D014760:Viral Fusion Proteins