title: Human cytomegalovirus final envelopment on membranes containing both trans-Golgi network and endosomal markers.
abstract: The human cytomegalovirus (HCMV) has been shown to complete its final envelopment on cytoplasmic membranes prior to its secretion to the extracellular medium. However, the nature of these membranes has not been characterized. It is thought that HCMV acquires its final envelope from the trans-Golgi network (TGN), though we and others have previously reported a role for endocytic membranes. Here we studied the localization of cellular markers in HCMV-infected cells and in isolated viruses. Immunofluorescence staining indicated that HCMV induces the recruitment of TGN and endosomal markers to the virus factory. Immuno-gold labelling of isolated viral particles and electron microscopy demonstrated the incorporation of TGN46, endosomal markers early endosomal antigen 1, annexin I, transferrin receptor and CD63, and the cation-independent mannose 6-phosphate receptor, which traffics between the TGN and endosomes into the viral envelope. Virus immunoprecipitation assays demonstrated that virions containing TGN46 and CD63 were infectious. This study reconciles the apparent controversy regarding the nature of the HCMV assembly site and suggests that HCMV has the ability to generate a novel membrane compartment containing markers for both TGN and endosomes, or that the membranes that HCMV uses for its envelope may be vesicles in transit between the TGN and endosomes.
authors.affiliation: Cell Biology of Herpesvirus Laboratory, Department of Molecular and Cell Biology, Centro Nacional de Biotecnología, Consejo Superior de Investigaciones Científicas, Campus Universidad Autonoma, Madrid 28049, Spain.
authors.forename: Victoria
authors.initials: V
authors.lastname: Cepeda
authors.forename: Mariano
authors.initials: M
authors.lastname: Esteban
authors.forename: Alberto
authors.initials: A
authors.lastname: Fraile-Ramos
mesh_terms: D002478:Cells, Cultured; D003587:Cytomegalovirus / Q000737:chemistry / Q000502:physiology*; D011992:Endosomes / Q000821:virology*; D005347:Fibroblasts / Q000821:virology; D006801:Humans; D047468:Immunoprecipitation; D007425:Intracellular Membranes / Q000737:chemistry; D008565:Membrane Proteins / Q000032:analysis; D008856:Microscopy, Fluorescence; D016253:Microscopy, Immunoelectron; D014763:Viral Matrix Proteins / Q000032:analysis*; D014771:Virion / Q000737:chemistry*; D019065:Virus Assembly; D021601:trans-Golgi Network / Q000821:virology*
chemical_list: D008565:Membrane Proteins; D014763:Viral Matrix Proteins