title: Role of cell surface spikes in alphavirus budding.
abstract: Alphaviruses mature by budding at cell surfaces. According to a prevailing hypothesis, the viral membrane protein, which is a heterodimeric protein unit, is transported to the plasma membrane (PM), where it awaits binding to the viral nucleocapsid (NC). This hypothesis predicts that the viral membrane protein heterodimers accumulate at the cell surface when expressed in the absence of NCs. We have tested this prediction by analyzing the spike protein expression phenotype of a Semliki Forest virus (SFV) variant which contains a capsid gene deletion. We found that viral membrane protein heterodimers were formed and transported to the cell surface normally. However, instead of accumulating at the PM as expected, the membrane proteins were rapidly degraded. In the case of the E1 subunit, degradation resulted in the release of a soluble E1 fragment into the medium. The fact that this pathway of protein degradation is mostly inhibited during wild-type virus infection suggests that viral membrane proteins are very efficiently captured by NCs into budding complexes and that normally no sizeable pool of free membrane protein complexes exists at the PM.
authors.affiliation: Department of Molecular Biology, Karolinska Institute, Huddinge, Sweden.
authors.forename: H
authors.initials: H
authors.lastname: Zhao
authors.forename: H
authors.initials: H
authors.lastname: Garoff
mesh_terms: D000524:Alphavirus / Q000254:growth & development / Q000502:physiology*; D000818:Animals; D002460:Cell Line; D002462:Cell Membrane / Q000502:physiology*; D006224:Cricetinae; D004705:Endocytosis; D017353:Gene Deletion; D005814:Genes, Viral; D007700:Kinetics; D046911:Macromolecular Substances; D008715:Methionine / Q000378:metabolism; D011499:Protein Processing, Post-Translational; D012672:Semliki forest virus / Q000235:genetics / Q000254:growth & development / Q000502:physiology*; D013462:Sulfur Radioisotopes; D014759:Viral Envelope Proteins / Q000096:biosynthesis* / Q000235:genetics; D014779:Virus Replication
chemical_list: D046911:Macromolecular Substances; D013462:Sulfur Radioisotopes; D014759:Viral Envelope Proteins; D008715:Methionine