1. Direct evidence that the GPCR CysLTR2 mutant causative of uveal melanoma is constitutively active with highly biased signaling
- Author
-
Jordan M. Mattheisen, Amanda R. Moore, Yu Chen, Ping Chi, Manija A. Kazmi, Tyler D. Hitchman, Mizuho Horioka, Thomas P. Sakmar, Emilie Ceraudo, and Thomas Huber
- Subjects
Uveal Neoplasms ,0301 basic medicine ,Receptors, Vasopressin ,Glutamine ,MV, missense variant ,Mutant ,HTRF, homogeneous time-resolved fluorescence immunoassay ,UVM, uveal melanoma ,Biochemistry ,Malignant transformation ,CysLTR2 ,Receptor ,Melanoma ,β-arrestins ,biased signaling ,CA, constitutive activity ,BRET, bioluminescent resonance energy transfer ,LTD4, leukotriene D4 ,beta-Arrestin 2 ,CAM, constitutively active mutant ,Cell biology ,Gene Expression Regulation, Neoplastic ,EPAC, exchange protein activated by cAMP ,LiCl, lithium chloride ,uveal melanoma ,Signal transduction ,signaling ,Protein Binding ,Signal Transduction ,Research Article ,PLCβ, phospholipase C-β ,Recombinant Fusion Proteins ,Green Fluorescent Proteins ,V2(A)6, hexa-alanine variant ,Biology ,Models, Biological ,RLuc3, Renilla Luciferase ,DPBS, Dulbecco’s PBS ,03 medical and health sciences ,FBS, fetal bovine serum ,Downregulation and upregulation ,Arrestin ,Humans ,BRET2, bioluminescent resonance energy transfer 2 ,constitutive activity ,Molecular Biology ,G protein-coupled receptor ,Receptors, Leukotriene ,CysLTR2, cysteinyl-leukotriene receptor 2 ,030102 biochemistry & molecular biology ,Lysine ,G protein ,Cell Biology ,CI, confidence interval ,Kinetics ,HEK293 Cells ,030104 developmental biology ,Cysteinyl leukotriene receptor 2 ,Amino Acid Substitution ,G protein–coupled receptor ,Mutation ,GTP-Binding Protein alpha Subunits, Gq-G11 ,GPCR, G protein–coupled receptor ,IP1, d-myo-inositol-1-phosphate ,BSA, bovine serum albumin - Abstract
Uveal melanoma is the most common eye cancer in adults and is clinically and genetically distinct from skin cutaneous melanoma. In a subset of cases, the oncogenic driver is an activating mutation in CYSLTR2, the gene encoding the G protein–coupled receptor cysteinyl-leukotriene receptor 2 (CysLTR2). The mutant CYSLTR2 encodes for the CysLTR2–L129Q receptor, with the substitution of Leu to Gln at position 129 (3.43). The ability of CysLTR2–L129Q to cause malignant transformation has been hypothesized to result from constitutive activity, but how the receptor could escape desensitization is unknown. Here, we characterize the functional properties of CysLTR2–L129Q. We show that CysLTR2–L129Q is a constitutively active mutant that strongly drives Gq/11 signaling pathways. However, CysLTR2–L129Q only poorly recruits β-arrestin. Using a modified Slack–Hall operational model, we quantified the constitutive activity for both pathways and conclude that CysLTR2–L129Q displays profound signaling bias for Gq/11 signaling pathways while escaping β-arrestin–mediated downregulation. CYSLTR2 is the first known example of a G protein–coupled receptor driver oncogene that encodes a highly biased constitutively active mutant receptor. These results provide new insights into the mechanism of CysLTR2–L129Q oncoprotein signaling and suggest CYSLTR2 as a promising potential therapeutic target in uveal melanoma.
- Published
- 2021