This Article
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrowReprints and Permissions
Right arrow Copyright Information
Right arrow Books from ASM Press
Right arrow MicrobeWorld
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Cohen, G H
Right arrow Articles by Eisenberg, R J
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Cohen, G H
Right arrow Articles by Eisenberg, R J

 Previous Article  |  Next Article 

J Virol. 1988 June; 62(6): 1932-1940

Expression of herpes simplex virus type 1 glycoprotein D deletion mutants in mammalian cells.

G H Cohen, W C Wilcox, D L Sodora, D Long, J Z Levin and R J Eisenberg

Department of Microbiology, School of Dental Medicine, University of Pennsylvania, Philadelphia 19104-6003.

ABSTRACT

Glycoprotein D (gD) is a viron envelope component of herpes simplex virus types 1 and 2. We have previously defined seven monoclonal antibody (MAb) groups which recognize distinct epitopes on the mature gD-1 protein of 369 amino acids. MAb groups VII, II, and V recognize continuous epitopes at residues 11-19, 272-279, and 340-356, respectively. MAb groups I, III, IV, and VI recognize discontinuous epitopes. Recent studies have focused on epitopes I, III, and VI. Using truncated forms of gD generated by recombinant DNA methods and proteolysis, epitopes III, IV, and VI were located within amino acids 1-233. A portion of discontinuous epitope I was located in a region within residues 233-275. For this study, we used recombinant DNA methods to create mutations in the gD-1 gene and studied the effects of those mutations on gD as expressed in mammalian cells. Plasmid pRE4, containing the coding sequence of gD-1 and the Rous sarcoma virus long terminal repeat promoter, was transfected into mammalian cells. The expressed protein, gD-1-(pRE4), was identical in size and antigenic properties to gD-1 from infected cells. Six in-frame deletion mutations were subsequently constructed by using restriction enzymes to excise portions of the gD-1 gene. Plasmids carrying these mutated forms were transfected into cells, and the corresponding proteins were examined at 48 h posttransfection for antigenicity and glycosylation patterns. Three deletions of varying size were located downstream of residue 233. Analysis of these mutants showed that amino acids within the region 234-244 were critical for binding of DL11 (group I), but not for other MAb groups. Three other deletion mutants lost all ability to bind MAbs which recognize discontinuous epitopes. In addition, much of the gD expressed by these mutants was observed to migrate as high-molecular-weight aggregated forms in nondenaturing gels. Each of these mutations involved the loss of a cysteine residue, suggesting that disulfide linkages play an essential role in the formation of discontinuous epitopes. The extent of glycosylation of the mutant gD molecules accumulated at 48 h posttransfection suggested altered carbohydrate processing. In one case, there was evidence for increased O-linked glycosylation. Those proteins which had lost a cysteine residue as part of the deletion did not accumulate molecules processed beyond the high-mannose stage. The results suggest that carbohydrate processing during synthesis of gD is very sensitive to alterations in structure, particularly changes involving cysteine residues.

J Virol. 1988 June; 62(6): 1932-1940




This article has been cited by other articles:

  • Mayer, D. C. G., Cofie, J., Jiang, L., Hartl, D. L., Tracy, E., Kabat, J., Mendoza, L. H., Miller, L. H. (2009). Glycophorin B is the erythrocyte receptor of Plasmodium falciparum erythrocyte-binding ligand, EBL-1. Proc. Natl. Acad. Sci. USA 106: 5348-5352 [Abstract] [Full Text]  
  • Ghumra, A., Semblat, J.-P., McIntosh, R. S., Raza, A., Rasmussen, I. B., Braathen, R., Johansen, F.-E., Sandlie, I., Mongini, P. K., Rowe, J. A., Pleass, R. J. (2008). Identification of Residues in the C{micro}4 Domain of Polymeric IgM Essential for Interaction with Plasmodium falciparum Erythrocyte Membrane Protein 1 (PfEMP1). J. Immunol. 181: 1988-2000 [Abstract] [Full Text]  
  • Bir, N., Yazdani, S. S., Avril, M., Layez, C., Gysin, J., Chitnis, C. E. (2006). Immunogenicity of Duffy Binding-Like Domains That Bind Chondroitin Sulfate A and Protection against Pregnancy-Associated Malaria.. Infect. Immun. 74: 5955-5963 [Abstract] [Full Text]  
  • Mayor, A., Bir, N., Sawhney, R., Singh, S., Pattnaik, P., Singh, S. K., Sharma, A., Chitnis, C. E. (2005). Receptor-binding residues lie in central regions of Duffy-binding-like domains involved in red cell invasion and cytoadherence by malaria parasites. Blood 105: 2557-2563 [Abstract] [Full Text]  
  • Lasaro, M. O., Luiz, W. B., Sbrogio-Almeida, M. E., Nishimura, L. S., Guth, B. E. C., Ferreira, L. C. S. (2004). Combined Vaccine Regimen Based on Parenteral Priming with a DNA Vaccine and Administration of an Oral Booster Consisting of a Recombinant Salmonella enterica Serovar Typhimurium Vaccine Strain for Immunization against Infection with Human-Derived Enterotoxigenic Escherichia coli Strains. Infect. Immun. 72: 6480-6491 [Abstract] [Full Text]  
  • Cocchi, F., Menotti, L., Di Ninni, V., Lopez, M., Campadelli-Fiume, G. (2004). The Herpes Simplex Virus JMP Mutant Enters Receptor-Negative J Cells through a Novel Pathway Independent of the Known Receptors nectin1, HveA, and nectin2. J. Virol. 78: 4720-4729 [Abstract] [Full Text]  
  • Mayer, D. C. G., Mu, J.-B., Kaneko, O., Duan, J., Su, X.-z., Miller, L. H. (2004). Polymorphism in the Plasmodium falciparum erythrocyte-binding ligand JESEBL/EBA-181 alters its receptor specificity. Proc. Natl. Acad. Sci. USA 101: 2518-2523 [Abstract] [Full Text]  
  • Milne, R. S. B., Hanna, S. L., Rux, A. H., Willis, S. H., Cohen, G. H., Eisenberg, R. J. (2003). Function of Herpes Simplex Virus Type 1 gD Mutants with Different Receptor-Binding Affinities in Virus Entry and Fusion. J. Virol. 77: 8962-8972 [Abstract] [Full Text]  
  • Chattopadhyay, R., Rathore, D., Fujioka, H., Kumar, S., de la Vega, P., Haynes, D., Moch, K., Fryauff, D., Wang, R., Carucci, D. J., Hoffman, S. L. (2003). PfSPATR, a Plasmodium falciparum Protein Containing an Altered Thrombospondin Type I Repeat Domain Is Expressed at Several Stages of the Parasite Life Cycle and Is the Target of Inhibitory Antibodies. J. Biol. Chem. 278: 25977-25981 [Abstract] [Full Text]  
  • Shukla, D., Dal Canto, M. C., Rowe, C. L., Spear, P. G. (2000). Striking Similarity of Murine Nectin-1alpha to Human Nectin-1alpha (HveC) in Sequence and Activity as a Glycoprotein D Receptor for Alphaherpesvirus Entry. J. Virol. 74: 11773-11781 [Abstract] [Full Text]  
  • Chen, Q., Heddini, A., Barragan, A., Fernandez, V., Pearce, S. F. A., Wahlgren, M. (2000). The Semiconserved Head Structure of Plasmodium falciparum Erythrocyte Membrane Protein 1 Mediates Binding to Multiple Independent Host Receptors. JEM 192: 1-10 [Abstract] [Full Text]  
  • Michon, P., Fraser, T., Adams, J. H. (2000). Naturally Acquired and Vaccine-Elicited Antibodies Block Erythrocyte Cytoadherence of the Plasmodium vivax Duffy Binding Protein. Infect. Immun. 68: 3164-3171 [Abstract] [Full Text]  
  • Whitbeck, J. C., Muggeridge, M. I., Rux, A. H., Hou, W., Krummenacher, C., Lou, H., van Geelen, A., Eisenberg, R. J., Cohen, G. H. (1999). The Major Neutralizing Antigenic Site on Herpes Simplex Virus Glycoprotein D Overlaps a Receptor-Binding Domain. J. Virol. 73: 9879-9890 [Abstract] [Full Text]  
  • Ranjan, A., Chitnis, C. E. (1999). Mapping regions containing binding residues within functional domains of Plasmodium vivax and Plasmodium knowlesi erythrocyte-binding proteins. Proc. Natl. Acad. Sci. USA 96: 14067-14072 [Abstract] [Full Text]  
  • Shukla, D., Rowe, C. L., Dong, Y., Racaniello, V. R., Spear, P. G. (1999). The Murine Homolog (Mph) of Human Herpesvirus Entry Protein B (HveB) Mediates Entry of Pseudorabies Virus but Not Herpes Simplex Virus Types 1 and 2. J. Virol. 73: 4493-4497 [Abstract] [Full Text]  
  • Rux, A. H., Willis, S. H., Nicola, A. V., Hou, W., Peng, C., Lou, H., Cohen, G. H., Eisenberg, R. J. (1998). Functional Region IV of Glycoprotein D from Herpes Simplex Virus Modulates Glycoprotein Binding to the Herpesvirus Entry Mediator. J. Virol. 72: 7091-7098 [Abstract] [Full Text]  
  • Kappe, S. H. I., Noe, A. R., Fraser, T. S., Blair, P. L., Adams, J. H. (1998). A family of chimeric erythrocyte binding proteins of malaria parasites. Proc. Natl. Acad. Sci. USA 95: 1230-1235 [Abstract] [Full Text]  
  • Sim, B., Chitnis, C., Wasniowska, K, Hadley, T., Miller, L. (1994). Receptor and ligand domains for invasion of erythrocytes by Plasmodium falciparum. Science 264: 1941-1944 [Abstract]  
  • Beghdadi-Rais, C, Schreyer, M, Rousseaux, M, Borel, P, Eisenberg, R., Cohen, G., Bron, C, Fasel, N (1993). Carboxyl terminus structural requirements for glycosyl-phosphatidylinositol anchor addition to cell surface proteins. J. Cell Sci. 105: 831-840 [Abstract]  
  • Singh, S., Pandey, K., Chattopadhayay, R., Yazdani, S. S., Lynn, A., Bharadwaj, A., Ranjan, A., Chitnis, C. (2001). Biochemical, Biophysical, and Functional Characterization of Bacterially Expressed and Refolded Receptor Binding Domain of Plasmodium vivax Duffy-binding Protein. J. Biol. Chem. 276: 17111-17116 [Abstract] [Full Text]  


Copyright © 2009 by the American Society for Microbiology.   For an alternate route to Journals.ASM.org, visit: http://intl-journals.asm.org | More Info»