title: The transmembrane domain sequence affects the structure and function of the Newcastle disease virus fusion protein.
abstract: The role of specific sequences in the transmembrane (TM) domain of Newcastle disease virus (NDV) fusion (F) protein in the structure and function of this protein was assessed by replacing this domain with the F protein TM domains from two other paramyxoviruses, Sendai virus (SV) and measles virus (MV), or the TM domain of the unrelated glycoprotein (G) of vesicular stomatitis virus (VSV). Mutant proteins with the SV or MV F protein TM domains were expressed, transported to cell surfaces, and proteolytically cleaved at levels comparable to that of the wild-type protein, while mutant proteins with the VSV G protein TM domain were less efficiently expressed on cell surfaces and proteolytically cleaved. All mutant proteins were defective in all steps of membrane fusion, including hemifusion. In contrast to the wild-type protein, the mutant proteins did not form detectable complexes with the NDV hemagglutinin-neuraminidase (HN) protein. As determined by binding of conformation-sensitive antibodies, the conformations of the ectodomains of the mutant proteins were altered. These results show that the specific sequence of the TM domain of the NDV F protein is important for the conformation of the preactivation form of the ectodomain, the interactions of the protein with HN protein, and fusion activity.
authors.affiliation: Department of Microbiology and Physiological Systems/Program in Immunology and Virology, University of Massachusetts Medical School, Worcester, MA 01655, USA.
authors.forename: Kathryn A
authors.initials: KA
authors.lastname: Gravel
authors.forename: Lori W
authors.initials: LW
authors.lastname: McGinnes
authors.forename: Julie
authors.initials: J
authors.lastname: Reitter
authors.forename: Trudy G
authors.initials: TG
authors.lastname: Morrison
mesh_terms: D000818:Animals; D019556:COS Cells; D002522:Chlorocebus aethiops; D015870:Gene Expression; D015707:HN Protein / Q000378:metabolism; D008459:Measles virus / Q000235:genetics; D050505:Mutant Proteins / Q000235:genetics / Q000378:metabolism; D009522:Newcastle disease virus / Q000737:chemistry / Q000235:genetics / Q000502:physiology*; D010447:Peptide Hydrolases / Q000378:metabolism; D011485:Protein Binding; D011487:Protein Conformation; D011995:Recombination, Genetic; D029082:Sendai virus / Q000235:genetics; D018116:Vesiculovirus / Q000235:genetics; D014760:Viral Fusion Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D053586:Virus Internalization
chemical_list: D015707:HN Protein; D050505:Mutant Proteins; D014760:Viral Fusion Proteins; D010447:Peptide Hydrolases