Identification and characterization of human PEIG-1/GPRC5A as a 12-Otetradecanoyl phorbol-13-acetate (TPA) and PKC-induced gene
| Autor: | Tomás A. Santa Coloma, María Á. Aguilar, Ángel G. Valdivieso, Consuelo Mori, María M. Massip-Copiz, Eduardo G. Cafferata, Mariángeles Clauzure |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2020 |
| Předmět: |
Protein Conformation
alpha-Helical 0301 basic medicine Indoles GPRC5A Biophysics PEIG-1 Biochemistry RAID-1 12 o tetradecanoyl phorbol 13 acetate Receptors G-Protein-Coupled Maleimides 03 medical and health sciences Cell Line Tumor Nitriles Butadienes Humans Amino Acid Sequence RNA Messenger PKC Receptor Egtazic Acid Molecular Biology Gene Protein Kinase C Protein kinase C Sulfonamides Differential display integumentary system 030102 biochemistry & molecular biology Chemistry Isoquinolines Molecular biology Protein Structure Tertiary Up-Regulation TIG1 030104 developmental biology Homo sapiens GenBank Tetradecanoylphorbol Acetate TPA RAI3 Signal Transduction |
| Zdroj: | Archives of Biochemistry and Biophysics Vol. 687,108375, 2020 Repositorio Institucional (UCA) Pontificia Universidad Católica Argentina instacron:UCA |
| Popis: | Fil: Mori, Consuelo. Pontificia Universidad Católica Argentina. Facultad de Ciencias Médicas. Instituto de Investigaciones Biomédicas. Laboratorio de Biología Celular y Molecular; Argentina Fil: Mori, Consuelo. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Valdivieso, Ángel Gabriel. Pontificia Universidad Católica Argentina. Facultad de Ciencias Médicas. Instituto de Investigaciones Biomédicas. Laboratorio de Biología Celular y Molecular; Argentina Fil: Valdivieso, Ángel Gabriel. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Clauzure, Mariángeles. Pontificia Universidad Católica Argentina. Facultad de Ciencias Médicas. Instituto de Investigaciones Biomédicas. Laboratorio de Biología Celular y Molecular; Argentina Fil: Clauzure, Mariángeles. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Massip Copiz, María M. Pontificia Universidad Católica Argentina. Facultad de Ciencias Médicas. Instituto de Investigaciones Biomédicas. Laboratorio de Biología Celular y Molecular; Argentina Fil: Massip Copiz, María M. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Aguilar, María Á. Pontificia Universidad Católica Argentina. Facultad de Ciencias Médicas. Instituto de Investigaciones Biomédicas. Laboratorio de Biología Celular y Molecular; Argentina Fil: Aguilar, María Á. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Cafferata, Eduardo G.A. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigaciones Bioquímicas de Buenos Aires. Fundación Instituto Leloir; Argentina Fil: Santa Coloma, Tomás Antonio. Pontificia Universidad Católica Argentina. Facultad de Ciencias Médicas. Instituto de Investigaciones Biomédicas. Laboratorio de Biología Celular y Molecular; Argentina Fil: Santa Coloma, Tomás Antonio. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Abstract: Homo sapiens orphan G protein-coupling receptor PEIG-1 was first cloned and characterized by applying differential display to T84 colonic carcinoma cells incubated in the presence of phorbol ester 12-O-tetradecanoylphorbol-13-acetate (TPA) (GenBank AF506289.1). Later, Lotan's laboratory found the same gene product in response to retinoic acid analogues, naming it with the symbol RAIG1. Now the official HGNC symbol is GPRC5A. Here, we report the extension of its original cDNA fragment towards the 5' and 3' end. In addition, we show that TPA (100 ng/ml, 162 nM) strongly stimulated GPRC5A mRNA in T84 colonic carcinoma cells, with maximal expression at 4 h and 100 ng/ml (162 nM). Western blots showed several bands between 35 and 50 kDa, responding to TPA stimulation. Confocal microscopy confirmed its TPA upregulation and the location in the plasma membrane. The PKC inhibitor Gö 6983 (10 μM), and the Ca2+ chelator BAPTA-AM (150 μM), strongly inhibited its TPA induced upregulation. The PKA inhibitor H-89 (10 μM), and the MEK1/2 inhibitor U0126 (10 μM), also produced a significant reduction in the TPA response (~50%). The SGK1 inhibitor GSK650394 stimulated GPRC5A basal levels at low doses and inhibit its TPA-induced expression at concentrations ≥10 μM. The IL-1β autocrine loop and downstream signalling did not affect its expression. In conclusion, RAIG1/RAI3/GPRC5A corresponds to the originally reported PEIG-1/TIG1; the inhibition observed in the presence of Gö 6983, BAPTA and U0126, suggests that its TPA-induced upregulation is mediated through a PKC/Ca2+ →MEK1/2 signalling axis. PKA and SGK1 kinases are also involved in its TPA-induced upregulation. |
| Databáze: | OpenAIRE |
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