title: Role of the transmembrane domain of marburg virus surface protein GP in assembly of the viral envelope.
abstract: The major protein constituents of the filoviral envelope are the matrix protein VP40 and the surface transmembrane protein GP. While VP40 is recruited to the sites of budding via the late retrograde endosomal transport route, GP is suggested to be transported via the classical secretory pathway involving the endoplasmic reticulum, Golgi apparatus, and trans-Golgi network until it reaches the plasma membrane where most filoviral budding takes place. Since both transport routes target the plasma membrane, it was thought that GP and VP40 join there to form the viral envelope. However, it was recently shown that, upon coexpression of both proteins, GP is partially recruited into peripheral VP40-enriched multivesicular bodies, which contained markers of the late endosome. Accumulation of GP and VP40 in this compartment was presumed to play an important role in the formation of the filoviral envelope. Using a domain-swapping approach, we were able to show that the transmembrane domain of GP was essential and sufficient for (i) partial recruitment of chimeric glycoproteins into VP40-enriched multivesicular bodies and (ii) incorporation into virus-like particles (VLPs) that were released upon expression of VP40. Only those chimeric glycoproteins which were targeted to VP40-enriched endosomal multivesicular bodies were subsequently recruited into VLPs. These data show that the transmembrane domain of GP is critical for the mixing of VP40 and GP in multivesicular bodies and incorporation of GP into the viral envelope. Results further suggest that trapping of GP in the VP40-enriched late endosomal compartment is important for the formation of the viral envelope.
authors.affiliation: Institute of Virology, Philipps University Marburg, and Robert Koch-Institut, Nordufer 20, 13353 Berlin, Germany.
authors.forename: Eva
authors.initials: E
authors.lastname: Mittler
authors.forename: Larissa
authors.initials: L
authors.lastname: Kolesnikova
authors.forename: Thomas
authors.initials: T
authors.lastname: Strecker
authors.forename: Wolfgang
authors.initials: W
authors.lastname: Garten
authors.forename: Stephan
authors.initials: S
authors.lastname: Becker
mesh_terms: D002460:Cell Line; D011992:Endosomes / Q000737:chemistry; D019084:Fluorescent Antibody Technique, Indirect; D006801:Humans; D029024:Marburgvirus / Q000502:physiology*; D008856:Microscopy, Fluorescence; D016253:Microscopy, Immunoelectron; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D014759:Viral Envelope Proteins / Q000235:genetics / Q000378:metabolism*; D022701:Virosomes / Q000378:metabolism / Q000648:ultrastructure; D019065:Virus Assembly
chemical_list: C081324:GP-protein, Marburg virus; D014759:Viral Envelope Proteins; D022701:Virosomes