Control of CXCR2 activity through its ubiquitination on K327 residue

BMC Cell Biology

Julie Gavard²  Nicolas Bidère¹  Julie Dwyer¹  Sandy Azzi¹  Sonia M Dubois¹  Héloise M Leclair¹ 

[1] Universite Paris Descartes, Sorbonne Paris Cite, 12 rue de l’Ecole de Medecine, Paris, 75006, France;Institut Cochin, 22 rue Mechain, Rm. 306, Paris, 75014, France

关键词: Signaling;    Internalization;    Arrestin;    Ubiquitination;    Chemokine receptor;   

Others  :  1088720
DOI  :  10.1186/s12860-014-0038-0

 received in 2014-06-26, accepted in 2014-10-09,  发布年份 2014

【 摘 要 】

Background

The interleukin-8 chemokine (IL-8) G-protein coupled receptor CXCR2 governs pro-inflammatory and pro-angiogenic responses in leukocytes and endothelial cells. At a molecular standpoint, CXCR2 is widely reported to operate through calcium flux, phosphoinoisitide 3 kinase (PI3K) and mitogen-activated protein kinase (MAPK). While CXCR2 trafficking is suspected to be intertwined with its signaling, the exact mechanism is not fully elucidated.

Results

Here, we identified the lysine 327 within the CXCR2 C-terminal tail as a key residue for ubiquitination, internalization, and signaling. First, the substitution to an arginine of K327 mutation was associated with a reduction in CXCR2 poly-ubiquitination. While WT CXCR2 was rapidly internalized following IL-8 administration, K327R mutant remained at the plasma membrane. Finally, K327R mutant failed to promote the recruitment of β-arrestin2, as estimated by imagery and bioluminescence resonance transfer. As a consequence, the activation of intracellular signaling, including both early events such as ERK phosphorylation and the increase in calcium flux, and the latter activation of the AP1 and NF-κB transcription factors, was blunted.

Conclusions

Overall, our results demonstrate that CXCR2 ubiquitination on K327 residue modulates agonist-activated CXCR2 cell sorting and intracellular signaling. Thus, the inhibition of K327 ubiquitination might emerge as an effective mean to curb exacerbated CXCR2 signaling in several pathological conditions, such as inflammatory diseases and cancer.

【 授权许可】

   
2014 Leclair et al.; licensee BioMed Central Ltd.

【 图 表 】

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【 参考文献 】

• [1]Addison CL, Daniel TO, Burdick MD, Liu H, Ehlert JE, Xue YY, Buechi L, Walz A, Richmond A, Strieter RM: The CXC chemokine receptor 2, CXCR2, is the putative receptor for ELR + CXC chemokine-induced angiogenic activity. J Immunol 2000, 165(9):5269-5277.
• [2]Dwyer J, Hebda JK, Le Guelte A, Galan-Moya EM, Smith SS, Azzi S, Bidere N, Gavard J: Glioblastoma cell-secreted interleukin-8 induces brain endothelial cell permeability via CXCR2. PLoS ONE 2012, 7(9):e45562.
• [3]Gavard J, Hou X, Qu Y, Masedunskas A, Martin D, Weigert R, Li X, Gutkind JS: A role for a CXCR2/phosphatidylinositol 3-kinase gamma signaling axis in acute and chronic vascular permeability. Mol Cell Biol 2009, 29(9):2469-2480.
• [4]Reutershan J: CXCR2–the receptor to hit? Drug News Perspect 2006, 19(10):615-623.
• [5]Koch AE, Polverini PJ, Kunkel SL, Harlow LA, DiPietro LA, Elner VM, Elner SG, Strieter RM: Interleukin-8 as a macrophage-derived mediator of angiogenesis. Science 1992, 258(5089):1798-1801.
• [6]Reutershan J, Morris MA, Burcin TL, Smith DF, Chang D, Saprito MS, Ley K: Critical role of endothelial CXCR2 in LPS-induced neutrophil migration into the lung. J Clin Invest 2006, 116(3):695-702.
• [7]Lazennec G, Richmond A: Chemokines and chemokine receptors: new insights into cancer-related inflammation. Trends Mol Med 2010, 16(3):133-144.
• [8]Ijichi H, Chytil A, Gorska AE, Aakre ME, Bierie B, Tada M, Mohri D, Miyabayashi K, Asaoka Y, Maeda S, Ikenoue T, Tateishi K, Wright CV, Koike K, Omata M, Moses HL: Inhibiting Cxcr2 disrupts tumor-stromal interactions and improves survival in a mouse model of pancreatic ductal adenocarcinoma. J Clin Invest 2011, 121(10):4106-4117.
• [9]Waugh DJJ, Wilson C: The Interleukin-8 Pathway in Cancer. Clin Cancer Res 2008, 14(21):6735-6741.
• [10]Cataisson C, Ohman R, Patel G, Pearson A, Tsien M, Jay S, Wright L, Hennings H, Yuspa SH: Inducible cutaneous inflammation reveals a protumorigenic role for keratinocyte CXCR2 in skin carcinogenesis. Cancer Res 2009, 69(1):319-328.
• [11]Dorsam RT, Gutkind JS: G-protein-coupled receptors and cancer. Nat Rev Cancer 2007, 7(2):79-94.
• [12]Marchese A, Trejo J: Ubiquitin-dependent regulation of G protein-coupled receptor trafficking and signaling. Cell Signal 2013, 25(3):707-716.
• [13]Baugher PJ, Richmond A: The carboxyl-terminal PDZ ligand motif of chemokine receptor CXCR2 modulates post-endocytic sorting and cellular chemotaxis. J Biol Chem 2008, 283(45):30868-30878.
• [14]Fan GH, Lapierre LA, Goldenring JR, Richmond A: Differential regulation of CXCR2 trafficking by Rab GTPases. Blood 2003, 101(6):2115-2124.
• [15]Azzi S, Smith SS, Dwyer J, Leclair HM, Alexia C, Hebda JK, Dupin N, Bidere N, Gavard J: YGLF motif in the Kaposi sarcoma herpes virus G-protein-coupled receptor adjusts NF-kappaB activation and paracrine actions. Oncogene 2013.
• [16]Fan GH, Yang W, Wang XJ, Qian Q, Richmond A: Identification of a motif in the carboxyl terminus of CXCR2 that is involved in adaptin 2 binding and receptor internalization. Biochemistry 2001, 40(3):791-800.
• [17]Neel NF, Barzik M, Raman D, Sobolik-Delmaire T, Sai J, Ham AJ, Mernaugh RL, Gertler FB, Richmond A: VASP is a CXCR2-interacting protein that regulates CXCR2-mediated polarization and chemotaxis. J Cell Sci 2009, 122(Pt 11):1882-1894.
• [18]Raman D, Sai J, Neel NF, Chew CS, Richmond A: LIM and SH3 protein-1 modulates CXCR2-mediated cell migration. PLoS ONE 2010, 5(4):e10050.
• [19]Raman D, Sai J, Hawkins O, Richmond A: Adaptor protein2 (AP2) orchestrates CXCR2-mediated cell migration. Traffic 2014, 15(4):451-469.
• [20]Dubois SM, Alexia C, Wu Y, Leclair HM, Leveau C, Schol E, Fest T, Tarte K, Chen ZJ, Gavard J, Bidere N: A catalytic-independent role for the LUBAC in NF-kappaB activation upon antigen receptor engagement and in lymphoma cells. Blood 2014, 123(14):2199-2203.
• [21]Oeckinghaus A, Wegener E, Welteke V, Ferch U, Arslan SC, Ruland J, Scheidereit C, Krappmann D: Malt1 ubiquitination triggers NF-kappaB signaling upon T-cell activation. EMBO J 2007, 26(22):4634-4645.
• [22]White JR, Lee JM, Young PR, Hertzberg RP, Jurewicz AJ, Chaikin MA, Widdowson K, Foley JJ, Martin LD, Griswold DE, Sarau HM: Identification of a potent, selective non-peptide CXCR2 antagonist that inhibits interleukin-8-induced neutrophil migration. J Biol Chem 1998, 273(17):10095-10098.
• [23]Lefkowitz RJ: Historical review: a brief history and personal retrospective of seven-transmembrane receptors. Trends Pharmacol Sci 2004, 25(8):413-422.
• [24]Hebda JK, Leclair HM, Azzi S, Roussel C, Scott MG, Bidere N, Gavard J: The C-terminus region of beta-arrestin1 modulates VE-cadherin expression and endothelial cell permeability. Cell Commun Signal 2013, 11(1):37. BioMed Central Full Text
• [25]Rappoport JZ, Taha BW, Lemeer S, Benmerah A, Simon SM: The AP-2 complex is excluded from the dynamic population of plasma membrane-associated clathrin. J Biol Chem 2003, 278(48):47357-47360.
• [26]Lakadamyali M, Rust MJ, Zhuang X: Ligands for clathrin-mediated endocytosis are differentially sorted into distinct populations of early endosomes. Cell 2006, 124(5):997-1009.
• [27]Wu S, Zhang Y: LOMETS: a local meta-threading-server for protein structure prediction. Nucleic Acids Res 2007, 35(10):3375-3382.
• [28]Gavard J, Gutkind JS: Protein kinase C-related kinase and ROCK are required for thrombin-induced endothelial cell permeability downstream from Galpha12/13 and Galpha11/q. J Biol Chem 2008, 283(44):29888-29896.
• [29]Charest PG, Terrillon S, Bouvier M: Monitoring agonist-promoted conformational changes of beta-arrestin in living cells by intramolecular BRET. EMBO Rep 2005, 6(4):334-340.