title: The conserved glycine residues in the transmembrane domain of the Semliki Forest virus fusion protein are not required for assembly and fusion.
abstract: The alphavirus Semliki Forest virus (SFV) infects cells via a low pH-triggered fusion reaction mediated by the viral E1 protein. Both the E1 fusion peptide and transmembrane (TM) domain are essential for membrane fusion, but the functional requirements for the TM domain are poorly understood. Here we explored the role of the five TM domain glycine residues, including the highly conserved glycine pair at E1 residues 415/416. SFV mutants with alanine substitutions for individual or all five glycine residues (5G/A) showed growth kinetics and fusion pH dependence similar to those of wild-type SFV. Mutants with increasing substitution of glycine residues showed an increasingly more stringent requirement for cholesterol during fusion. The 5G/A mutant showed decreased fusion kinetics and extent in fluorescent lipid mixing assays. TM domain glycine residues thus are not required for efficient SFV fusion or assembly but can cause subtle effects on the properties of membrane fusion.
authors.affiliation: Department of Cell Biology, Albert Einstein College of Medicine, 1300 Morris Park Avenue, Bronx, NY 10461, USA.
authors.forename: Maofu
authors.initials: M
authors.lastname: Liao
authors.forename: Margaret
authors.initials: M
authors.lastname: Kielian
mesh_terms: D000818:Animals; D002460:Cell Line; D017124:Conserved Sequence; D006224:Cricetinae; D005998:Glycine / Q000235:genetics / Q000502:physiology*; D008561:Membrane Fusion / Q000502:physiology*; D008565:Membrane Proteins / Q000502:physiology; D009154:Mutation; D010455:Peptides / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D017434:Protein Structure, Tertiary / Q000502:physiology; D012672:Semliki forest virus / Q000502:physiology*; D014158:Transcription, Genetic; D014760:Viral Fusion Proteins / Q000235:genetics / Q000502:physiology*; D019065:Virus Assembly
chemical_list: D008565:Membrane Proteins; D010455:Peptides; D014760:Viral Fusion Proteins; D005998:Glycine