Summary
Surface biotinylation of human cytomegalovirus (HCMV)-infected fibroblasts under pulse-chase conditions was used to define the cellular route of the dominant viral envelope glycoprotein gB into the cytoplasmic compartment of viral maturational envelopment. The results showed that a major fraction of gB was re-internalized from the infected cell surface prior to incorporation into the viral envelope. Viral particles carrying biotinylated gB were subsequently released into the culture medium. Viral release appeared to be inhibited in the presence of gB-specific antibody or when infected cultures were incubated at room temperature, but was not reduced by inhibitors of cellular glycoprotein transport. To our knowledge this is the first report describing that HCMV gB is retrieved from the infected cell surface prior to viral envelopment.
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References
Benko D, Gibson W (1989) A major tegument phosphoprotein of human cytomegalovirus contains O-linked GlcNAc. In: Compans RW, Helenius A, Oldstone MB (eds) Cell biology of virus entry, replication and pathogenesis. Alan R. Liss New York, pp 57–66
Bogner R, Reschke M, Reis B, Reis E, Britt W, Radsak K (1992) Recognition of compartmentalized intracellular analogs of glycoprotein H of human cytomegalovirus. Arch Virol 126: 67–80
Britt W, Vugler L (1987) Structural and immunological characterization of the intracellular forms of an abundant 68000 Mr human cytomegalovirus protein. J Gen Virol 68: 1897–1907
Campadelli-fiume G, Farabegoli F, Digaeta S, Roizman B (1991) Origin of unenveloped capsids in the cytoplasm of cells infected with herpes simplex virus 1. J Virol 65: 1589–1595
Carter L, Redelmeier T, Wollenweber L, Schmid S (1993) Multiple GTP-binding proteins participate in clathrin-coated vesicle-mediated endocytosis. J Cell Biol 120: 37–45
Chapman R, Munro S (1994) Retrieval of TGN proteins from the cell surface requires endosomal acidification. EMBO J 13: 2305–2312
Cranage M, Smith G, Bell S, Hart H, Brown C, Bankier A, Tomlinson P, Barrell B, Minson T (1988) Identification and expression of a human cytomegalovirus glycoprotein with homology to the Epstein-Barr virus BXLF2 product, varicella-zoster virus gpIII, and herpes simplex virus type 1 glycoprotein H. J Virol 62: 1416–1422
Cheung P, Banfield B, Tufaro S (1991) Brefeldin A arrests the maturation and egress of herpes simplex virus particles during infection. J Virol 65: 1893–1904
Collawn JF, Lai A, Domingo D, Fitch M, Hatton S, Trowbridge IS (1993) YTRF is the conserved internalization signal of the transferrin receptor, and a second YTRF signal at position 31–34 enhances endocytosis. J Biol Chem 268: 21686–21692
Eggers M, Bogner E, Agricola B, Kern H, Radsak K (1992) Inhibition of human cytomegalovirus maturation by brefeldin A. J Gen Virol 73: 2679–2692
Eis-Hübinger A, Schmidt D, Schneweis K (1993) Anti-glycoprotein B monoclonal antibody protects T cell depleted mice against herpes simplex virus infection by inhibition of virus replication at the inoculated mucous membranes. J Gen Virol 74: 379–385
Gabel C, Dubey L, Steinberg S, Sherman D, Gershon M, Gershon A (1989) Varicellazoster virus glycoprotein oligosaccharides are phosphorylated during posttranslational maturation. J Virol 63: 4264–4276
Gershon A, Sherman D, Zhu Z, Gabel C, Ambron R, Gershon M (1994) Intracellular transport of newly synthesized varizella-zoster virus: final envelopment in the trans-Golgi network. J Virol 68: 6372–6390
Griffiths G, Hoflack B, Simons K, Mellman I, Kornfeld S (1988) The mannose 6-phosphate receptor and the biogenesis of lysosomes. Cell 52: 239–341
Iwayama S, Yamamoto T, Furuya T, Kobayashi R, Ikuta K, Hirai K (1994) Intracellular localization and DNA binding activity of a class of viral early phosphoproteins in human fibroblasts infected with human cytomegalovirus (Towne strain). J Gen Virol 75: 3309–3318
Kaiser C, Radsak K (1987) Inhibition by monensin of human cytomegalovirus DNA replication. Arch Virol 94: 229–245
Kühn J, Kramer M, Willenbacher W, Wieland U, Lorentzen E, Braun R (1990) Identification of herpes simplex virus type 1 glycoproteins interacting with the cell surface. J Virol 64: 2491–2497
Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227: 680–685
Ludwig T, Griffiths G, Hoflack B (1991) Distribution of newly synthesized lysosomal enzymes in the endocytic pathway of normal rat kidney cells. J Cell Biol 115: 1561–1572
Luzio JP, Banting G (1993) Eukaryotic membrane traffic: retrieval and retention mechanisms to achieve organelle residence. Trends Biochem Sci 18: 395–398
Ohlin M, Sunquist V, Mach M, Wahren B, Borrebaeck C (1993) Fine specificity of the human immune response to the major neutralizing epitopes expressed on cytomegalovirus gp58/116 (gB), as determined with human monoclonal antibodies. J Virol 67: 703–710
Radsak K, Schneider D, Jost E, Brücher K (1989) Alteration of nuclear lamina protein in human fibroblasts infected with cytomegalovirus. Arch Virol 105: 103–112
Radsak K, Brücher K, Britt W, Shiou H, Schneider D, Kollert A (1990) Nuclear compartmentation of glycoprotein B of human cytomegalovirus. Virology 177: 515–522
Radsak K, Brücher K, Georgatos S (1991) Focal nuclear lesions and specific nuclear lamin A/C dephosphorylation during infection with human cytomegalovirus. Eur J Cell Biol 54: 299–304
Reis B, Bogner E, Reschke M, Richter A, Mockenhaupt T, Radsak K (1993) Stable constitutive expression of glycoprotein B (gpUL55) of human cytomegalovirus in permissive astrocytoma cells. J Gen Virol 74: 1371–1379
Reschke M, Reis B, Nöding K, Rohsiepe D, Richter A, Mockenhaupt T, Garten W, Radsak K (1995) Constitutive expression of human cytomegalovirus glycoprotein B (gpUL55) with mutagenized carboxyterminal hydrophobic domains. J Gen Virol 76: 113–122
Sairenji T, Bertoni G, Medveczky M, Medveczky P, Nguyen Q, Humphreys R (1988) Inhibition of Epstein-Barr virus (EBV) release from P3HR-1 and B95-8 lines by monoclonal antibodies to EBV membrane antigen gp350/220. J Virol 62: 2614–2521
Schmelz M, Sodeik B, Ericsson M, Wolffe E, Shida H, Hiller G, Griffiths G (1994) Assembly of vaccinia virus: the second wrapping cisterna is derived from the trans Golgi network. J Virol 68: 130–147
Severi B, Landini M, Govoni E (1988) Human cytomegalovirus morphogenesis: an ultrastructural study of the late cytoplasmic phases. Arch Virol 98: 51–64
Spaete R, Thayer R, Probert W, Masiarz F, Chamberlain S, Rasmussen L, Merigan T, Pachl C (1988) Human cytomegalovirus strain towne glycoprotein B is processed by proteolytic cleavage. Virology 167: 207–225
Tooze J, Hollinshead M, Reis B, Radsak K, Kern H (1993) Progeny vaccinia virus and human cytomegalovirus particles utilize early endosomal cisternae for their envelopment. Eur J Cell Biol 60: 163–178
Van Deurs B, Sandvig K, Petersen O, Olsnes S, Simons K, Griffiths G (1988) Estimation of the amount of internalized ricin that reaches the trans-Golgi network. J Cell Biol 106: 253–267
Whealy M, Card J, Meade R, Robbins A, Enquist L (1991) Effect of brefeldin A on alpha-herpesvirus membrane protein glycosylation and virus egress. J Virol 65: 1066–1081
Wood S, Park J, Brown W (1991) Brefeldin A causes a microtubule-mediated fusion of the trans-Golgi network and early endosomes. Cell 67: 591–600
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Radsak, K., Eickmann, M., Mockenhaupt, T. et al. Retrieval of human cytomegalovirus glycoprotein B from the infected cell surface for virus envelopment. Archives of Virology 141, 557–572 (1996). https://doi.org/10.1007/BF01718317
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DOI: https://doi.org/10.1007/BF01718317