title: A single amino acid change in E3 of ts1 mutant inhibits the intracellular transport of SFV envelope protein complex.
abstract: At 39 degrees the envelope protein complex (E1-p62) of Semliki Forest virus mutant ts1 is arrested in the rough endoplasmic reticulum (RER). When the infected cultures are shifted to 28 degrees, the complex is transported to the cell surface. During the transport p62 is cleaved into E2 under conditions in which no virus budding takes place. We have sequenced the cDNA, which encodes the envelope proteins of ts1. Comparison with the respective wild-type nucleotide sequence shows only one nucleotide change, G----A, causing a replacement of cysteine-58 (TGC) with tyrosine (TAC) in the E3 protein of ts1. A cDNA fragment from the ts1 genome encoding the mutation in E3 was used to replace the respective fragment of prototype SFV in an eukaryotic expression vector. Intracellular arrest of envelope proteins at 39 degrees was seen in transfected BHK21 cells. A shift of the transfected cells to 28 degrees resulted in the appearance of the envelope proteins at the cell surface. We conclude that the single point mutation is solely responsible for the temperature-sensitive transport defect of ts1 envelope glycoproteins.
authors.affiliation: Institute of Biotechnology, University of Helsinki, Finland.
authors.forename: P
authors.initials: P
authors.lastname: Syväoja
authors.forename: J
authors.initials: J
authors.lastname: Peränen
authors.forename: M
authors.initials: M
authors.lastname: Suomalainen
authors.forename: S
authors.initials: S
authors.lastname: Keränen
authors.forename: L
authors.initials: L
authors.lastname: Kääriäinen
mesh_terms: D001483:Base Sequence; D002451:Cell Compartmentation; D003001:Cloning, Molecular; D004247:DNA / Q000235:genetics; D005455:Fluorescent Antibody Technique; D005822:Genetic Vectors; D008969:Molecular Sequence Data; D009154:Mutation; D009841:Oligonucleotides; D011499:Protein Processing, Post-Translational; D015183:Restriction Mapping; D012672:Semliki forest virus / Q000235:genetics / Q000378:metabolism*; D013329:Structure-Activity Relationship; D014759:Viral Envelope Proteins / Q000235:genetics / Q000378:metabolism*
chemical_list: D009841:Oligonucleotides; D014759:Viral Envelope Proteins; D004247:DNA