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 Minisini, R.
Right arrow Articles by Moepps, B.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Minisini, R.
Right arrow Articles by Moepps, B.

 Previous Article  |  Next Article 

Journal of Virology, April 2003, p. 4489-4501, Vol. 77, No. 8
0022-538X/03/$08.00+0     DOI: 10.1128/JVI.77.8.4489-4501.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Constitutive Inositol Phosphate Formation in Cytomegalovirus-Infected Human Fibroblasts Is due to Expression of the Chemokine Receptor Homologue pUS28

Rosalba Minisini,1 Calogero Tulone,2 Anke Lüske,1 Detlef Michel,1 Thomas Mertens,1 Peter Gierschik,2* and Barbara Moepps2

Departments of Virology,1 Pharmacology and Toxicology, University of Ulm, 89081 Ulm, Germany2

Received 14 August 2002/ Accepted 14 January 2003

An open reading frame (ORF), US28, with homology to mammalian chemokine receptors has been identified in the genome of human cytomegalovirus (HCMV). Its protein product, pUS28, has been shown to bind several human CC chemokines, including RANTES, MCP-1, and MIP-1{alpha}, and the CX3C chemokine fractalkine with high affinity. Addition of CC chemokines to cells expressing pUS28 was reported to cause a pertussis toxin-sensitive increase in the concentration of cytosolic free Ca2+. Recently, pUS28 was shown to mediate constitutive, ligand-independent, and pertussis toxin-insensitive activation of phospholipase C via Gq/11-dependent signaling pathways in transiently transfected COS-7 cells. Since these findings are not easily reconciled with the former observations, we analyzed the role of pUS28 in mediating CC chemokine activation of pertussis toxin-sensitive G proteins in cell membranes and phospholipase C in intact cells. The transmembrane signaling functions of pUS28 were studied in HCMV-infected cells rather than in cDNA-transfected cells. Since DNA sequence analysis of ORF US28 of different laboratory and clinical strains had revealed amino acid sequence differences in the amino-terminal portion of pUS28, we compared two laboratory HCMV strains, AD169 and Toledo, and one clinical strain, TB40/E. The results showed that infection of human fibroblasts with all three HCMV strains led to a vigorous, constitutively enhanced formation of inositol phosphates which was insensitive to pertussis toxin. This effect was critically dependent on the presence of the US28 ORF in the HCMV genome but was independent of the amino acid sequence divergence of the three HCMV strains investigated. The constitutive activity of pUS28 is not explained by expression of pUS28 at high density in HCMV-infected cells. The pUS28 ligands RANTES and MCP-1 failed to stimulate binding of guanosine 5'-O-(3-[35S]thiotriphosphate to membranes of HCMV-infected cells and did not enhance constitutive activation of phospholipase C in intact HCMV-infected cells. These findings raise the possibility that the effects of CC chemokines and pertussis toxin on G protein-mediated transmembrane signaling previously observed in HCMV-infected cells are either independent of or not directly mediated by the protein product of ORF US28.

* Corresponding author. Mailing address: Department of Pharmacology and Toxicology, University of Ulm, Albert-Einstein-Allee 11, 89081 Ulm, Germany. Phone: 49-731-5002 3870. Fax: 49-731-5002 3872. E-mail: peter.gierschik{at}medizin.uni-ulm.de.

Journal of Virology, April 2003, p. 4489-4501, Vol. 77, No. 8
0022-538X/03/$08.00+0     DOI: 10.1128/JVI.77.8.4489-4501.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Stropes, M. P., Schneider, O. D., Zagorski, W. A., Miller, J. L. C., Miller, W. E. (2009). The Carboxy-Terminal Tail of Human Cytomegalovirus (HCMV) US28 Regulates both Chemokine-Independent and Chemokine-Dependent Signaling in HCMV-Infected Cells. J. Virol. 83: 10016-10027 [Abstract] [Full Text]  
  • Stropes, M. P. M., Miller, W. E. (2008). Functional analysis of human cytomegalovirus pUS28 mutants in infected cells. J. Gen. Virol. 89: 97-105 [Abstract] [Full Text]  
  • Maussang, D., Verzijl, D., van Walsum, M., Leurs, R., Holl, J., Pleskoff, O., Michel, D., van Dongen, G. A. M. S., Smit, M. J. (2006). Human cytomegalovirus-encoded chemokine receptor US28 promotes tumorigenesis. Proc. Natl. Acad. Sci. USA 103: 13068-13073 [Abstract] [Full Text]  
  • Fitzsimons, C. P., Gompels, U. A., Verzijl, D., Vischer, H. F., Mattick, C., Leurs, R., Smit, M. J. (2006). Chemokine-Directed Trafficking of Receptor Stimulus to Different G Proteins: Selective Inducible and Constitutive Signaling by Human Herpesvirus 6-Encoded Chemokine Receptor U51. Mol. Pharmacol. 69: 888-898 [Abstract] [Full Text]  
  • Casarosa, P., Waldhoer, M., LiWang, P. J., Vischer, H. F., Kledal, T., Timmerman, H., Schwartz, T. W., Smit, M. J., Leurs, R. (2005). CC and CX3C Chemokines Differentially Interact with the N Terminus of the Human Cytomegalovirus-encoded US28 Receptor. J. Biol. Chem. 280: 3275-3285 [Abstract] [Full Text]  
  • Michel, D., Milotic, I., Wagner, M., Vaida, B., Holl, J., Ansorge, R., Mertens, T. (2005). The human cytomegalovirus UL78 gene is highly conserved among clinical isolates, but is dispensable for replication in fibroblasts and a renal artery organ-culture system. J. Gen. Virol. 86: 297-306 [Abstract] [Full Text]  
  • Reinhardt, B., Schaarschmidt, P., Bossert, A., Luske, A., Finkenzeller, G., Mertens, T., Michel, D. (2005). Upregulation of functionally active vascular endothelial growth factor by human cytomegalovirus. J. Gen. Virol. 86: 23-30 [Abstract] [Full Text]  
  • Weber, M., Blair, E., Simpson, C. V., O'Hara, M., Blackburn, P. E., Rot, A., Graham, G. J., Nibbs, R. J.B. (2004). The Chemokine Receptor D6 Constitutively Traffics to and from the Cell Surface to Internalize and Degrade Chemokines. Mol. Biol. Cell 15: 2492-2508 [Abstract] [Full Text]  
  • Casarosa, P., Gruijthuijsen, Y. K., Michel, D., Beisser, P. S., Holl, J., Fitzsimons, C. P., Verzijl, D., Bruggeman, C. A., Mertens, T., Leurs, R., Vink, C., Smit, M. J. (2003). Constitutive Signaling of the Human Cytomegalovirus-encoded Receptor UL33 Differs from That of Its Rat Cytomegalovirus Homolog R33 by Promiscuous Activation of G Proteins of the Gq, Gi, and Gs Classes. J. Biol. Chem. 278: 50010-50023 [Abstract] [Full Text]  
  • Streblow, D. N., Vomaske, J., Smith, P., Melnychuk, R., Hall, L., Pancheva, D., Smit, M., Casarosa, P., Schlaepfer, D. D., Nelson, J. A. (2003). Human Cytomegalovirus Chemokine Receptor US28-induced Smooth Muscle Cell Migration Is Mediated by Focal Adhesion Kinase and Src. J. Biol. Chem. 278: 50456-50465 [Abstract] [Full Text]  
  • Penfold, M. E. T., Schmidt, T. L., Dairaghi, D. J., Barry, P. A., Schall, T. J. (2003). Characterization of the Rhesus Cytomegalovirus US28 Locus. J. Virol. 77: 10404-10413 [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»