This Article
Right arrow Full Text
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 Strive, T.
Right arrow Articles by Radsak, K.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Strive, T.
Right arrow Articles by Radsak, K.

 Previous Article  |  Next Article 

Journal of Virology, February 2002, p. 1252-1264, Vol. 76, No. 3
0022-538X/01/$04.00+0     DOI: 10.1128/JVI.76.3.1252-1264.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Proteolytic Processing of Human Cytomegalovirus Glycoprotein B Is Dispensable for Viral Growth in Culture

Tanja Strive,1 Eva Borst,2 Martin Messerle,2 and Klaus Radsak1*

Institut für Virologie der Philipps-Universität, 35037 Marburg,1 Max-von-Pettenkofer-Institut, Lehrstuhl für Virologie, Genzentrum, Ludwig-Maximilians-Universität, 81377 Munich, Germany2

Received 25 May 2001/ Accepted 25 October 2001

Glycoprotein B (gB) of human cytomegalovirus (HCMV), which is considered essential for the viral life cycle, is proteolytically processed during maturation. Since gB homologues of several other herpesviruses remain uncleaved, the relevance of this property of HCMV gB for viral infectivity is unclear. Here we report on the construction of a viral mutant in which the recognition site of gB for the cellular endoprotease furin was destroyed. Because mutagenesis of essential proteins may result in a lethal phenotype, a replication-deficient HCMV gB-null genome encoding enhanced green fluorescent protein was constructed, and complementation by mutant gBs was initially evaluated in transient-cotransfection assays. Cotransfection of plasmids expressing authentic gB or gB with a mutated cleavage site (gB-{Delta}Fur) led to the formation of green fluorescent miniplaques which were considered to result from one cycle of phenotypic complementation of the gB-null genome. To verify these results, two recombinant HCMV genomes were constructed: HCMV-BAC-{Delta}MhdI, with a deletion of hydrophobic domain 1 of gB that appeared to be essential for viral growth in the cotransfection experiments, and HCMV-BAC{Delta}Fur, in which the gB cleavage site was mutated by amino acid substitution. Consistent with the results of the cotransfection assays, only the {Delta}Fur mutant replicated in human fibroblasts, showing growth kinetics comparable to that of wild-type virus. gB in mutant-infected cells was uncleaved, whereas glycosylation and transport to the cell surface were not impaired. Extracellular mutant virus contained exclusively uncleaved gB, indicating that proteolytic processing of gB is dispensable for viral replication in cell culture.

* Corresponding author. Mailing address: Institut für Virologie der Philipps-Universität, Robert-Koch-Strasse 17, 35037 Marburg, Germany. Phone: 49 (0)6421 28-64315. Fax: 49 (0)6421 28-63154. E-mail: radsak{at}mailer.uni-marburg.de.

Journal of Virology, February 2002, p. 1252-1264, Vol. 76, No. 3
0022-538X/01/$04.00+0     DOI: 10.1128/JVI.76.3.1252-1264.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Oliver, S. L., Sommer, M., Zerboni, L., Rajamani, J., Grose, C., Arvin, A. M. (2009). Mutagenesis of Varicella-Zoster Virus Glycoprotein B: Putative Fusion Loop Residues Are Essential for Viral Replication, and the Furin Cleavage Motif Contributes to Pathogenesis in Skin Tissue In Vivo. J. Virol. 83: 7495-7506 [Abstract] [Full Text]  
  • Sorem, J., Longnecker, R. (2009). Cleavage of Epstein-Barr virus glycoprotein B is required for full function in cell-cell fusion with both epithelial and B cells. J. Gen. Virol. 90: 591-595 [Abstract] [Full Text]  
  • Reimer, J. J., Backovic, M., Deshpande, C. G., Jardetzky, T., Longnecker, R. (2009). Analysis of Epstein-Barr Virus Glycoprotein B Functional Domains via Linker Insertion Mutagenesis. J. Virol. 83: 734-747 [Abstract] [Full Text]  
  • Nagel, C.-H., Dohner, K., Fathollahy, M., Strive, T., Borst, E. M., Messerle, M., Sodeik, B. (2008). Nuclear Egress and Envelopment of Herpes Simplex Virus Capsids Analyzed with Dual-Color Fluorescence HSV1(17+). J. Virol. 82: 3109-3124 [Abstract] [Full Text]  
  • Bergeron, E., Vincent, M. J., Nichol, S. T. (2007). Crimean-Congo Hemorrhagic Fever Virus Glycoprotein Processing by the Endoprotease SKI-1/S1P Is Critical for Virus Infectivity. J. Virol. 81: 13271-13276 [Abstract] [Full Text]  
  • Okazaki, K. (2007). Proteolytic cleavage of glycoprotein B is dispensable for in vitro replication, but required for syncytium formation of pseudorabies virus. J. Gen. Virol. 88: 1859-1865 [Abstract] [Full Text]  
  • Mavinakere, M. S., Williamson, C. D., Goldmacher, V. S., Colberg-Poley, A. M. (2006). Processing of Human Cytomegalovirus UL37 Mutant Glycoproteins in the Endoplasmic Reticulum Lumen prior to Mitochondrial Importation. J. Virol. 80: 6771-6783 [Abstract] [Full Text]  
  • Li, W., Minova-Foster, T. J., Norton, D. D., Muggeridge, M. I. (2006). Identification of functional domains in herpes simplex virus 2 glycoprotein B.. J. Virol. 80: 3792-3800 [Abstract] [Full Text]  
  • Sampaio, K. L., Cavignac, Y., Stierhof, Y.-D., Sinzger, C. (2005). Human Cytomegalovirus Labeled with Green Fluorescent Protein for Live Analysis of Intracellular Particle Movements. J. Virol. 79: 2754-2767 [Abstract] [Full Text]  
  • Keil, G. M., Hohle, C., Giesow, K., Konig, P. (2005). Engineering Glycoprotein B of Bovine Herpesvirus 1 To Function as Transporter for Secreted Proteins: a New Protein Expression Approach. J. Virol. 79: 791-799 [Abstract] [Full Text]  
  • Hansen, S. G., Strelow, L. I., Franchi, D. C., Anders, D. G., Wong, S. W. (2003). Complete Sequence and Genomic Analysis of Rhesus Cytomegalovirus. J. Virol. 77: 6620-6636 [Abstract] [Full Text]  
  • Chang, W. L. W., Barry, P. A. (2003). Cloning of the Full-Length Rhesus Cytomegalovirus Genome as an Infectious and Self-Excisable Bacterial Artificial Chromosome for Analysis of Viral Pathogenesis. J. Virol. 77: 5073-5083 [Abstract] [Full Text]  
  • Zimmer, G., Conzelmann, K.-K., Herrler, G. (2002). Cleavage at the Furin Consensus Sequence RAR/KR109 and Presence of the Intervening Peptide of the Respiratory Syncytial Virus Fusion Protein Are Dispensable for Virus Replication in Cell Culture. J. Virol. 76: 9218-9224 [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»