Abstract
The disulfide-linked glycoprotein B (gB; gp55-116) complex of human cytomegalovirus represents the most abundant and immunogenic component of the virion envelope. We have studied the oligomerization and transport of this molecule, using a series of murine monoclonal antibodies. Our results indicated that oligomerization of this molecule occurred shortly after its synthesis, with a half-time of maximal formation of approximately 25 min. The oligomeric form had an estimated mass of 340,000 Da and likely consisted of a homodimer of the gp55-116 complex. By using a conformation-specific monoclonal antibody, postoligomerization folding of this molecule was demonstrated. This event exhibited an unusually prolonged half-maximal time of approximately 160 min. Both oligomerization and folding occurred in the endoplasmic reticulum. Oligomerization and folding occurred in the absence of carbohydrate modifications, although likely at lower efficiency. Finally, the oligomeric and folded forms were shown to be transported to the surface of infected cells and infectious virions.
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- Bowden R. A., Fisher L. D., Rogers K., Cays M., Meyers J. D. Cytomegalovirus (CMV)-specific intravenous immunoglobulin for the prevention of primary CMV infection and disease after marrow transplant. J Infect Dis. 1991 Sep;164(3):483–487. doi: 10.1093/infdis/164.3.483. [DOI] [PubMed] [Google Scholar]
- Braakman I., Helenius J., Helenius A. Role of ATP and disulphide bonds during protein folding in the endoplasmic reticulum. Nature. 1992 Mar 19;356(6366):260–262. doi: 10.1038/356260a0. [DOI] [PubMed] [Google Scholar]
- Britt W. J., Auger D. Synthesis and processing of the envelope gp55-116 complex of human cytomegalovirus. J Virol. 1986 Apr;58(1):185–191. doi: 10.1128/jvi.58.1.185-191.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Britt W. J. Neutralizing antibodies detect a disulfide-linked glycoprotein complex within the envelope of human cytomegalovirus. Virology. 1984 Jun;135(2):369–378. doi: 10.1016/0042-6822(84)90193-4. [DOI] [PubMed] [Google Scholar]
- Britt W. J. Recent advances in the identification of significant human cytomegalovirus-encoded proteins. Transplant Proc. 1991 Jun;23(3 Suppl 3):64-9, discussion 69. [PubMed] [Google Scholar]
- Britt W. J., Vugler L. G. Processing of the gp55-116 envelope glycoprotein complex (gB) of human cytomegalovirus. J Virol. 1989 Jan;63(1):403–410. doi: 10.1128/jvi.63.1.403-410.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Britt W. J., Vugler L., Butfiloski E. J., Stephens E. B. Cell surface expression of human cytomegalovirus (HCMV) gp55-116 (gB): use of HCMV-recombinant vaccinia virus-infected cells in analysis of the human neutralizing antibody response. J Virol. 1990 Mar;64(3):1079–1085. doi: 10.1128/jvi.64.3.1079-1085.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Dorner A. J., Wasley L. C., Kaufman R. J. Protein dissociation from GRP78 and secretion are blocked by depletion of cellular ATP levels. Proc Natl Acad Sci U S A. 1990 Oct;87(19):7429–7432. doi: 10.1073/pnas.87.19.7429. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Gething M. J., McCammon K., Sambrook J. Expression of wild-type and mutant forms of influenza hemagglutinin: the role of folding in intracellular transport. Cell. 1986 Sep 12;46(6):939–950. doi: 10.1016/0092-8674(86)90076-0. [DOI] [PubMed] [Google Scholar]
- Gething M. J., Sambrook J. Protein folding in the cell. Nature. 1992 Jan 2;355(6355):33–45. doi: 10.1038/355033a0. [DOI] [PubMed] [Google Scholar]
- Gretch D. R., Gehrz R. C., Stinski M. F. Characterization of a human cytomegalovirus glycoprotein complex (gcI). J Gen Virol. 1988 Jun;69(Pt 6):1205–1215. doi: 10.1099/0022-1317-69-6-1205. [DOI] [PubMed] [Google Scholar]
- Gretch D. R., Kari B., Rasmussen L., Gehrz R. C., Stinski M. F. Identification and characterization of three distinct families of glycoprotein complexes in the envelopes of human cytomegalovirus. J Virol. 1988 Mar;62(3):875–881. doi: 10.1128/jvi.62.3.875-881.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hurtley S. M., Helenius A. Protein oligomerization in the endoplasmic reticulum. Annu Rev Cell Biol. 1989;5:277–307. doi: 10.1146/annurev.cb.05.110189.001425. [DOI] [PubMed] [Google Scholar]
- Hutter J. A., Scott J., Wreghitt T., Higenbottam T., Wallwork J. The importance of cytomegalovirus in heart-lung transplant recipients. Chest. 1989 Mar;95(3):627–631. doi: 10.1378/chest.95.3.627. [DOI] [PubMed] [Google Scholar]
- Jacobson M. A., Mills J. Serious cytomegalovirus disease in the acquired immunodeficiency syndrome (AIDS). Clinical findings, diagnosis, and treatment. Ann Intern Med. 1988 Apr;108(4):585–594. doi: 10.7326/0003-4819-108-4-585. [DOI] [PubMed] [Google Scholar]
- Kari B., Liu Y. N., Goertz R., Lussenhop N., Stinski M. F., Gehrz R. Structure and composition of a family of human cytomegalovirus glycoprotein complexes designated gC-I (gB). J Gen Virol. 1990 Nov;71(Pt 11):2673–2680. doi: 10.1099/0022-1317-71-11-2673. [DOI] [PubMed] [Google Scholar]
- Kniess N., Mach M., Fay J., Britt W. J. Distribution of linear antigenic sites on glycoprotein gp55 of human cytomegalovirus. J Virol. 1991 Jan;65(1):138–146. doi: 10.1128/jvi.65.1.138-146.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Manservigi R., Grossi M. P., Gualandri R., Balboni P. G., Marchini A., Rotola A., Rimessi P., Di Luca D., Cassai E., Barbanti-Brodano G. Protection from herpes simplex virus type 1 lethal and latent infections by secreted recombinant glycoprotein B constitutively expressed in human cells with a BK virus episomal vector. J Virol. 1990 Jan;64(1):431–436. doi: 10.1128/jvi.64.1.431-436.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Meyers J. D., Flournoy N., Thomas E. D. Risk factors for cytomegalovirus infection after human marrow transplantation. J Infect Dis. 1986 Mar;153(3):478–488. doi: 10.1093/infdis/153.3.478. [DOI] [PubMed] [Google Scholar]
- Pereira L., Hoffman M., Gallo D., Cremer N. Monoclonal antibodies to human cytomegalovirus: three surface membrane proteins with unique immunological and electrophoretic properties specify cross-reactive determinants. Infect Immun. 1982 Jun;36(3):924–932. doi: 10.1128/iai.36.3.924-932.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Rose J. K., Doms R. W. Regulation of protein export from the endoplasmic reticulum. Annu Rev Cell Biol. 1988;4:257–288. doi: 10.1146/annurev.cb.04.110188.001353. [DOI] [PubMed] [Google Scholar]
- Singh N., Dummer J. S., Kusne S., Breinig M. K., Armstrong J. A., Makowka L., Starzl T. E., Ho M. Infections with cytomegalovirus and other herpesviruses in 121 liver transplant recipients: transmission by donated organ and the effect of OKT3 antibodies. J Infect Dis. 1988 Jul;158(1):124–131. doi: 10.1093/infdis/158.1.124. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Snydman D. R., Werner B. G., Heinze-Lacey B., Berardi V. P., Tilney N. L., Kirkman R. L., Milford E. L., Cho S. I., Bush H. L., Jr, Levey A. S. Use of cytomegalovirus immune globulin to prevent cytomegalovirus disease in renal-transplant recipients. N Engl J Med. 1987 Oct 22;317(17):1049–1054. doi: 10.1056/NEJM198710223171703. [DOI] [PubMed] [Google Scholar]
- Spaete R. R. A recombinant subunit vaccine approach to HCMV vaccine development. Transplant Proc. 1991 Jun;23(3 Suppl 3):90–96. [PubMed] [Google Scholar]
- Spaete R. R., Saxena A., Scott P. I., Song G. J., Probert W. S., Britt W. J., Gibson W., Rasmussen L., Pachl C. Sequence requirements for proteolytic processing of glycoprotein B of human cytomegalovirus strain Towne. J Virol. 1990 Jun;64(6):2922–2931. doi: 10.1128/jvi.64.6.2922-2931.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Urban M., Britt W., Mach M. The dominant linear neutralizing antibody-binding site of glycoprotein gp86 of human cytomegalovirus is strain specific. J Virol. 1992 Mar;66(3):1303–1311. doi: 10.1128/jvi.66.3.1303-1311.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Utz U., Britt W., Vugler L., Mach M. Identification of a neutralizing epitope on glycoprotein gp58 of human cytomegalovirus. J Virol. 1989 May;63(5):1995–2001. doi: 10.1128/jvi.63.5.1995-2001.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Whealy M. E., Robbins A. K., Enquist L. W. The export pathway of the pseudorabies virus gB homolog gII involves oligomer formation in the endoplasmic reticulum and protease processing in the Golgi apparatus. J Virol. 1990 May;64(5):1946–1955. doi: 10.1128/jvi.64.5.1946-1955.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]