title: Adverse effects of MVA-T7 on the transport of Marburg virus glycoprotein.
abstract: Expression of glycoproteins has been carried out successfully using recombinant vaccinia virus vectors. Especially attractive is the use of recombinant vaccinia viruses which express the DNA-dependent RNA polymerase of the phage T7 (T7-polymerase). The T7-polynerase drives the transcription of plasmid-based genes under the control of the T7 RNA polymerase promoter transfected into the infected cell. Comparison of two different recombinant vaccinia viruses, vTF7-3 and MVA-T7, revealed that post-translational processing of Marburg virus surface glycoprotein (GP) is impaired in the MVA-T7 but not in the vTF7-3 system. Influenza virus hemagglutinin, however, was transported and processed like the authentic protein in both systems. It is shown that transport of GP in the MVA-T7 system is not completely blocked, but the vast majority of molecules remained Endo H-sensitive. Only trace amounts evaded the endoplasmatic reticulum and reached the plasma membrane. Thus, the adverse effects of MVA-T7 on the processing of recombinant glycoproteins cannot be predicted, and correct processing has to be investigated for every expressed glycoprotein.
authors.affiliation: Institut für Virologie der Philipps, Universität Marburg, Robert-Koch-Strasse 17, D-35037, Marburg, Germany.
authors.forename: C
authors.initials: C
authors.lastname: Sänger
authors.forename: E
authors.initials: E
authors.lastname: Mühlberger
authors.forename: H D
authors.initials: HD
authors.lastname: Klenk
authors.forename: S
authors.initials: S
authors.lastname: Becker
mesh_terms: D002462:Cell Membrane / Q000378:metabolism; D004721:Endoplasmic Reticulum / Q000378:metabolism; D005822:Genetic Vectors; D006367:HeLa Cells; D006389:Hemagglutinins, Viral / Q000235:genetics / Q000378:metabolism; D006801:Humans; D009980:Influenza A virus / Q000235:genetics / Q000378:metabolism; D011499:Protein Processing, Post-Translational; D021381:Protein Transport; D014616:Vaccinia virus / Q000235:genetics*; D014759:Viral Envelope Proteins / Q000235:genetics / Q000378:metabolism*
chemical_list: C081324:GP-protein, Marburg virus; D006389:Hemagglutinins, Viral; D014759:Viral Envelope Proteins