Helix 8 of the viral chemokine receptor ORF74 directs chemokine binding

Dennis Verzijl, Leonardo Pardo, Marie Van Dijk, Yvonne K. Gruijthuijsen, Aldo Jongejan, Henk Timmerman, John Nicholas, Mario Schwarz, Philip M. Murphy, Rob Leurs, Martine J. Smit

Research output: Contribution to journalArticle

Abstract

The constitutively active G-protein-coupled receptor and viral oncogene ORF74, encoded by Kaposi sarcoma-associated herpesvirus (human herpesvirus 8), binds a broad range of chemokines, including CXCL1 (agonist), CXCL8 (neutral ligand), and CXCL10 (inverse agonist). Although chemokines interact with the extracellular N terminus and loops of the receptor, we demonstrate that helix 8 (Hx8) in the intracellular carboxyl tail (C-tail) of ORF74 directs chemokine binding. Partial deletion of the C-tail resulted in a phenotype with reduced constitutive activity but intact regulation by ligands. Complete deletion of the C-tail, including Hx8, resulted in an inactive phenotype that lacks CXCL8 binding sites and has an increased number of binding sites for CXCL10. Similar effects were obtained with the single R7.61322W or Q7.62 323P mutations in Hx8. We propose that the conserved charged or polar side chain at position 7.61 has a specific role in stabilizing the end of transmembrane domain 7 (TM7). Disruption of Hx8 by deletion or mutation distorts an H-bonding network, involving highly conserved amino acids within TM2, TM7, and Hx8, that is crucial for positioning of the TM domains, coupling to Gαq, and CXCL8 binding. Thus, Hx8 appears to exert a key role in receptor stabilization through the conserved residue R7.61, directing the ligand binding profile of ORF74 and likely also that of other class A G-protein-coupled receptors.

Original languageEnglish (US)
Pages (from-to)35327-35335
Number of pages9
JournalJournal of Biological Chemistry
Volume281
Issue number46
DOIs
StatePublished - Nov 17 2006

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Chemokine Receptors
Chemokines
Human Herpesvirus 8
G-Protein-Coupled Receptors
Ligands
Chemokine CXCL1
Binding Sites
Phenotype
Sequence Deletion
Oncogenes
Stabilization
Amino Acids
Mutation

ASJC Scopus subject areas

  • Biochemistry

Cite this

Verzijl, D., Pardo, L., Van Dijk, M., Gruijthuijsen, Y. K., Jongejan, A., Timmerman, H., ... Smit, M. J. (2006). Helix 8 of the viral chemokine receptor ORF74 directs chemokine binding. Journal of Biological Chemistry, 281(46), 35327-35335. https://doi.org/10.1074/jbc.M606877200

Helix 8 of the viral chemokine receptor ORF74 directs chemokine binding. / Verzijl, Dennis; Pardo, Leonardo; Van Dijk, Marie; Gruijthuijsen, Yvonne K.; Jongejan, Aldo; Timmerman, Henk; Nicholas, John; Schwarz, Mario; Murphy, Philip M.; Leurs, Rob; Smit, Martine J.

In: Journal of Biological Chemistry, Vol. 281, No. 46, 17.11.2006, p. 35327-35335.

Research output: Contribution to journalArticle

Verzijl, D, Pardo, L, Van Dijk, M, Gruijthuijsen, YK, Jongejan, A, Timmerman, H, Nicholas, J, Schwarz, M, Murphy, PM, Leurs, R & Smit, MJ 2006, 'Helix 8 of the viral chemokine receptor ORF74 directs chemokine binding', Journal of Biological Chemistry, vol. 281, no. 46, pp. 35327-35335. https://doi.org/10.1074/jbc.M606877200
Verzijl D, Pardo L, Van Dijk M, Gruijthuijsen YK, Jongejan A, Timmerman H et al. Helix 8 of the viral chemokine receptor ORF74 directs chemokine binding. Journal of Biological Chemistry. 2006 Nov 17;281(46):35327-35335. https://doi.org/10.1074/jbc.M606877200
Verzijl, Dennis ; Pardo, Leonardo ; Van Dijk, Marie ; Gruijthuijsen, Yvonne K. ; Jongejan, Aldo ; Timmerman, Henk ; Nicholas, John ; Schwarz, Mario ; Murphy, Philip M. ; Leurs, Rob ; Smit, Martine J. / Helix 8 of the viral chemokine receptor ORF74 directs chemokine binding. In: Journal of Biological Chemistry. 2006 ; Vol. 281, No. 46. pp. 35327-35335.
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