Mutation of the DRY motif reveals different structural requirements for the CC chemokine receptor 5-mediated signaling and receptor endocytosis. Export

@article{citeulike:356002, abstract = {CC chemokine receptor 5 (CCR5) is a G protein-coupled receptor that governs migration of leukocytes and serves as a coreceptor for the R5 tropic strains of human immunodeficiency virus (HIV). CCR5-mediated signaling in response to CC chemokines relies on G protein activation. Desensitization, which rapidly turns off G protein-dependent signaling, involves phosphorylation of CCR5 that promotes interaction of the receptor with beta-arrestins for endocytosis. Whether coupling to G proteins, desensitization, and endocytosis of CCR5 require the same structural determinants remains a matter of investigation. Here, we show that CCR5 displayed agonist-independent coupling to G proteins. This constitutive activity of the receptor was abrogated by TAK779 (N,N-dimethyl-N-[4-[[[2-(4-methylphenyl)-6,7-dihydro-5H-benzocyclohepten-8-yl]carbonyl]amino]benzyl]tetrahydro-2H-pyran-4-aminium chloride), a nonpeptidic CCR5 ligand that inhibits HIV infection and was found to depend on the integrity of the Asp-Arg-Tyr (DRY) motif. Changing Arg-126 by the neutral residue Asn (R126N-CCR5 mutant) abolished CCR5-mediated activation of G proteins, either constitutively or in response to agonists. In contrast, R126N-CCR5 not only retained agonist-promoted phosphorylation and beta-arrestin-dependent endocytosis but also displayed a higher basal phosphorylation than wild-type CCR5. Expression of beta-arrestin in R126N-CCR5-expressing cells resulted in receptor down-regulation, thereby suggesting that R126N-CCR5 spontaneously interacts with beta-arrestins. However, although expression of beta-arrestin favored wild-type CCR5-mediated chemotaxis, it failed to promote migration of cells expressing R126N-CCR5. Overall, these data indicate that structural requirements for CCR5-mediated activation of G proteins, albeit not involved in receptor desensitization and internalization, are needed for beta-arrestin-mediated chemotaxis. These results have implications for how distinct biological responses of CCR5 might rely on a different set of receptor conformations.}, address = {Institut Pasteur, Unit\'{e} d'Immunologie Virale, 28 rue du Dr Roux, 75015 Paris, France.}, author = {Lagane, B. and Ballet, S. and Planchenault, T. and Balabanian, K. and Le Poul, E. and Blanpain, C. and Percherancier, Y. and Staropoli, I. and Vassart, G. and Oppermann, M. and Parmentier, M. and Bachelerie, F.}, citeulike-article-id = {356002}, citeulike-linkout-0 = {http://dx.doi.org/10.1124/mol.104.009779}, citeulike-linkout-1 = {http://molpharm.aspetjournals.org/cgi/content/abstract/67/6/1966}, citeulike-linkout-2 = {http://view.ncbi.nlm.nih.gov/pubmed/15761117}, citeulike-linkout-3 = {http://www.hubmed.org/display.cgi?uids=15761117}, doi = {10.1124/mol.104.009779}, issn = {0026-895X}, journal = {Mol Pharmacol}, keywords = {ccr5, dry, oppermann, trafficking}, month = {June}, number = {6}, pages = {1966--1976}, posted-at = {2005-10-25 17:04:05}, priority = {0}, title = {Mutation of the DRY motif reveals different structural requirements for the CC chemokine receptor 5-mediated signaling and receptor endocytosis.}, url = {http://dx.doi.org/10.1124/mol.104.009779}, volume = {67}, year = {2005} }

TY - JOUR ID - citeulike:356002 L3 - citeulike-article-id:356002 N2 - CC chemokine receptor 5 (CCR5) is a G protein-coupled receptor that governs migration of leukocytes and serves as a coreceptor for the R5 tropic strains of human immunodeficiency virus (HIV). CCR5-mediated signaling in response to CC chemokines relies on G protein activation. Desensitization, which rapidly turns off G protein-dependent signaling, involves phosphorylation of CCR5 that promotes interaction of the receptor with beta-arrestins for endocytosis. Whether coupling to G proteins, desensitization, and endocytosis of CCR5 require the same structural determinants remains a matter of investigation. Here, we show that CCR5 displayed agonist-independent coupling to G proteins. This constitutive activity of the receptor was abrogated by TAK779 (N,N-dimethyl-N-[4-[[[2-(4-methylphenyl)-6,7-dihydro-5H-benzocyclohepten-8-yl]carbonyl]amino]benzyl]tetrahydro-2H-pyran-4-aminium chloride), a nonpeptidic CCR5 ligand that inhibits HIV infection and was found to depend on the integrity of the Asp-Arg-Tyr (DRY) motif. Changing Arg-126 by the neutral residue Asn (R126N-CCR5 mutant) abolished CCR5-mediated activation of G proteins, either constitutively or in response to agonists. In contrast, R126N-CCR5 not only retained agonist-promoted phosphorylation and beta-arrestin-dependent endocytosis but also displayed a higher basal phosphorylation than wild-type CCR5. Expression of beta-arrestin in R126N-CCR5-expressing cells resulted in receptor down-regulation, thereby suggesting that R126N-CCR5 spontaneously interacts with beta-arrestins. However, although expression of beta-arrestin favored wild-type CCR5-mediated chemotaxis, it failed to promote migration of cells expressing R126N-CCR5. Overall, these data indicate that structural requirements for CCR5-mediated activation of G proteins, albeit not involved in receptor desensitization and internalization, are needed for beta-arrestin-mediated chemotaxis. These results have implications for how distinct biological responses of CCR5 might rely on a different set of receptor conformations. IS - 6 JF - Mol Pharmacol SN - 0026-895X AD - Institut Pasteur, Unité d'Immunologie Virale, 28 rue du Dr Roux, 75015 Paris, France. EP - 1976 TI - Mutation of the DRY motif reveals different structural requirements for the CC chemokine receptor 5-mediated signaling and receptor endocytosis. VL - 67 SP - 1966 KW - ccr5 KW - dry KW - oppermann KW - trafficking AU - Lagane, B AU - Ballet, S AU - Planchenault, T AU - Balabanian, K AU - Le Poul, E AU - Blanpain, C AU - Percherancier, Y AU - Staropoli, I AU - Vassart, G AU - Oppermann, M AU - Parmentier, M AU - Bachelerie, F PY - 2005/06// UR - http://dx.doi.org/10.1124/mol.104.009779 ER -