title: The aromatic domain of the coronavirus class I viral fusion protein induces membrane permeabilization: putative role during viral entry.
abstract: Coronavirus (CoV) entry is mediated by the viral spike (S) glycoprotein, a class I viral fusion protein. During viral and target cell membrane fusion, the heptad repeat (HR) regions of the S2 subunit assume a trimer-of-hairpins structure, positioning the fusion peptide in close proximity to the C-terminal region of the ectodomain. The formation of this structure appears to drive apposition and subsequent fusion of viral and target cell membranes; however, the exact mechanism is unclear. Here, we characterize an aromatic amino acid rich region within the ectodomain of the S2 subunit that both partitions into lipid membranes and has the capacity to perturb lipid vesicle integrity. Circular dichroism analysis indicated that peptides analogous to the aromatic domains of the severe acute respiratory syndrome (SARS)-CoV, mouse hepatitis virus (MHV) and the human CoV OC43 S2 subunits, did not have a propensity for a defined secondary structure. These peptides strongly partitioned into lipid membranes and induced lipid vesicle permeabilization at peptide/lipid ratios of 1:100 in two independent leakage assays. Thus, partitioning of the peptides into the lipid interface is sufficient to disorganize membrane integrity. Our study of the S2 aromatic domain of three CoVs provides supportive evidence for a functional role of this region. We propose that, when aligned with the fusion peptide and transmembrane domains during membrane apposition, the aromatic domain of the CoV S protein functions to perturb the target cell membrane and provides a continuous track of hydrophobic surface, resulting in lipid-membrane fusion and subsequent viral nucleocapsid entry.
authors.affiliation: Department of Microbiology and Immunology, Program in Molecular Pathogenesis and Immunity, Tulane University Health Sciences Center, New Orleans, Louisiana 70112, USA.
authors.forename: Bruno
authors.initials: B
authors.lastname: Sainz
authors.forename: Joshua M
authors.initials: JM
authors.lastname: Rausch
authors.forename: William R
authors.initials: WR
authors.lastname: Gallaher
authors.forename: Robert F
authors.initials: RF
authors.lastname: Garry
authors.forename: William C
authors.initials: WC
authors.lastname: Wimley
mesh_terms: D000595:Amino Acid Sequence; D002942:Circular Dichroism; D017934:Coronavirus / Q000502:physiology*; D008561:Membrane Fusion / Q000502:physiology*; D008969:Molecular Sequence Data; D017433:Protein Structure, Secondary; D040901:Proteomics; D014760:Viral Fusion Proteins / Q000737:chemistry / Q000502:physiology*
chemical_list: D014760:Viral Fusion Proteins