Abstract
In this study, reverse transcriptase polymerase chain reaction was used to amplify human endothelin receptor A (ETA) and ETB receptor mRNA. A truncated ETA receptor transcript with exons 3 and 4 skipped was found. The skipping of these two exons results in 109 amino acids being deleted from the receptor. The truncated receptor was expressed in all tissues and cells examined, but the level of expression varied. In melanoma cell lines and melanoma tissues, the truncated receptor gene was the major species, whereas the wild-type ETA was predominant in other tissues. A 1.9-kb ETA transcript was identified in melanoma cell lines by Northern blot, which was much smaller than the transcript in heart and in other tissues reported previously (4.3 kb). The cDNA coding regions of the truncated and wild-type ETA receptors were stably transfected into Chinese hamster ovary (CHO) cells. The truncated ETA receptor-transfected CHO cells did not show binding affinity to endothelin 1 (ET-1) or endothelin 3 (ET-3). The function and biological significance of this truncated ETA receptor is not clear, but it may have regulatory roles for cell responses to ETs.
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- Adachi M., Furuichi Y., Miyamoto C. Identification of a ligand-binding site of the human endothelin-A receptor and specific regions required for ligand selectivity. Eur J Biochem. 1994 Feb 15;220(1):37–43. doi: 10.1111/j.1432-1033.1994.tb18596.x. [DOI] [PubMed] [Google Scholar]
- Arai H., Hori S., Aramori I., Ohkubo H., Nakanishi S. Cloning and expression of a cDNA encoding an endothelin receptor. Nature. 1990 Dec 20;348(6303):730–732. doi: 10.1038/348730a0. [DOI] [PubMed] [Google Scholar]
- Arai H., Nakao K., Takaya K., Hosoda K., Ogawa Y., Nakanishi S., Imura H. The human endothelin-B receptor gene. Structural organization and chromosomal assignment. J Biol Chem. 1993 Feb 15;268(5):3463–3470. [PubMed] [Google Scholar]
- Bax W. A., Bos E., Saxena P. R. Heterogeneity of endothelin/sarafotoxin receptors mediating contraction of the human isolated saphenous vein. Eur J Pharmacol. 1993 Aug 3;239(1-3):267–268. doi: 10.1016/0014-2999(93)91010-k. [DOI] [PubMed] [Google Scholar]
- Becker A., Haendler B., Hechler U., Schleuning W. D. Mutational analysis of human endothelin receptors ETA and ETB identification of regions involved in the selectivity for endothelin 3 or cyclo-(D-Trp-D-Asp-Pro-D-Val-Leu). Eur J Biochem. 1994 May 1;221(3):951–958. doi: 10.1111/j.1432-1033.1994.tb18810.x. [DOI] [PubMed] [Google Scholar]
- Canton H., Emeson R. B., Barker E. L., Backstrom J. R., Lu J. T., Chang M. S., Sanders-Bush E. Identification, molecular cloning, and distribution of a short variant of the 5-hydroxytryptamine2C receptor produced by alternative splicing. Mol Pharmacol. 1996 Oct;50(4):799–807. [PubMed] [Google Scholar]
- Chomczynski P., Sacchi N. Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal Biochem. 1987 Apr;162(1):156–159. doi: 10.1006/abio.1987.9999. [DOI] [PubMed] [Google Scholar]
- Elshourbagy N. A., Adamou J. E., Gagnon A. W., Wu H. L., Pullen M., Nambi P. Molecular characterization of a novel human endothelin receptor splice variant. J Biol Chem. 1996 Oct 11;271(41):25300–25307. doi: 10.1074/jbc.271.41.25300. [DOI] [PubMed] [Google Scholar]
- Faure E., Gouédard L., Imbeaud S., Cate R., Picard J. Y., Josso N., di Clemente N. Mutant isoforms of the anti-Müllerian hormone type II receptor are not expressed at the cell membrane. J Biol Chem. 1996 Nov 29;271(48):30571–30575. doi: 10.1074/jbc.271.48.30571. [DOI] [PubMed] [Google Scholar]
- Giros B., Martres M. P., Pilon C., Sokoloff P., Schwartz J. C. Shorter variants of the D3 dopamine receptor produced through various patterns of alternative splicing. Biochem Biophys Res Commun. 1991 May 15;176(3):1584–1592. doi: 10.1016/0006-291x(91)90469-n. [DOI] [PubMed] [Google Scholar]
- Hosoda K., Nakao K., Hiroshi-Arai, Suga S., Ogawa Y., Mukoyama M., Shirakami G., Saito Y., Nakanishi S., Imura H. Cloning and expression of human endothelin-1 receptor cDNA. FEBS Lett. 1991 Aug 5;287(1-2):23–26. doi: 10.1016/0014-5793(91)80007-p. [DOI] [PubMed] [Google Scholar]
- Hosoda K., Nakao K., Tamura N., Arai H., Ogawa Y., Suga S., Nakanishi S., Imura H. Organization, structure, chromosomal assignment, and expression of the gene encoding the human endothelin-A receptor. J Biol Chem. 1992 Sep 15;267(26):18797–18804. [PubMed] [Google Scholar]
- Inoue A., Yanagisawa M., Kimura S., Kasuya Y., Miyauchi T., Goto K., Masaki T. The human endothelin family: three structurally and pharmacologically distinct isopeptides predicted by three separate genes. Proc Natl Acad Sci U S A. 1989 Apr;86(8):2863–2867. doi: 10.1073/pnas.86.8.2863. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kikuchi K., Nakagawa H., Kadono T., Etoh T., Byers H. R., Mihm M. C., Tamaki K. Decreased ET(B) receptor expression in human metastatic melanoma cells. Biochem Biophys Res Commun. 1996 Feb 27;219(3):734–739. doi: 10.1006/bbrc.1996.0303. [DOI] [PubMed] [Google Scholar]
- Mombouli J. V., Le S. Q., Wasserstrum N., Vanhoutte P. M. Endothelins 1 and 3 and big endothelin-1 contract isolated human placental veins. J Cardiovasc Pharmacol. 1993;22 (Suppl 8):S278–S281. doi: 10.1097/00005344-199322008-00073. [DOI] [PubMed] [Google Scholar]
- Ogawa Y., Nakao K., Arai H., Nakagawa O., Hosoda K., Suga S., Nakanishi S., Imura H. Molecular cloning of a non-isopeptide-selective human endothelin receptor. Biochem Biophys Res Commun. 1991 Jul 15;178(1):248–255. doi: 10.1016/0006-291x(91)91806-n. [DOI] [PubMed] [Google Scholar]
- Reid K., Turnley A. M., Maxwell G. D., Kurihara Y., Kurihara H., Bartlett P. F., Murphy M. Multiple roles for endothelin in melanocyte development: regulation of progenitor number and stimulation of differentiation. Development. 1996 Dec;122(12):3911–3919. doi: 10.1242/dev.122.12.3911. [DOI] [PubMed] [Google Scholar]
- Riezebos J., Watts I. S., Vallance P. J. Endothelin receptors mediating functional responses in human small arteries and veins. Br J Pharmacol. 1994 Feb;111(2):609–615. doi: 10.1111/j.1476-5381.1994.tb14780.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Sakamoto A., Yanagisawa M., Sawamura T., Enoki T., Ohtani T., Sakurai T., Nakao K., Toyo-oka T., Masaki T. Distinct subdomains of human endothelin receptors determine their selectivity to endothelinA-selective antagonist and endothelinB-selective agonists. J Biol Chem. 1993 Apr 25;268(12):8547–8553. [PubMed] [Google Scholar]
- Sakurai T., Yanagisawa M., Takuwa Y., Miyazaki H., Kimura S., Goto K., Masaki T. Cloning of a cDNA encoding a non-isopeptide-selective subtype of the endothelin receptor. Nature. 1990 Dec 20;348(6303):732–735. doi: 10.1038/348732a0. [DOI] [PubMed] [Google Scholar]
- Schöneberg T., Liu J., Wess J. Plasma membrane localization and functional rescue of truncated forms of a G protein-coupled receptor. J Biol Chem. 1995 Jul 28;270(30):18000–18006. doi: 10.1074/jbc.270.30.18000. [DOI] [PubMed] [Google Scholar]
- Shyamala V., Moulthrop T. H., Stratton-Thomas J., Tekamp-Olson P. Two distinct human endothelin B receptors generated by alternative splicing from a single gene. Cell Mol Biol Res. 1994;40(4):285–296. [PubMed] [Google Scholar]
- Tso J. Y., Sun X. H., Kao T. H., Reece K. S., Wu R. Isolation and characterization of rat and human glyceraldehyde-3-phosphate dehydrogenase cDNAs: genomic complexity and molecular evolution of the gene. Nucleic Acids Res. 1985 Apr 11;13(7):2485–2502. doi: 10.1093/nar/13.7.2485. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Yohn J. J., Smith C., Stevens T., Hoffman T. A., Morelli J. G., Hurt D. L., Yanagisawa M., Kane M. A., Zamora M. R. Human melanoma cells express functional endothelin-1 receptors. Biochem Biophys Res Commun. 1994 May 30;201(1):449–457. doi: 10.1006/bbrc.1994.1722. [DOI] [PubMed] [Google Scholar]