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Regulation of GPCR expression through an interaction with CCT7, a subunit of the CCT/TRiC complex
Mechanisms that prevent aggregation and promote folding of nascent G protein–coupled receptors (GPCRs) remain poorly understood. We identified chaperonin containing TCP-1 subunit eta (CCT7) as an interacting partner of the β-isoform of thromboxane A(2) receptor (TPβ) by yeast two-hybrid screening. C...
Autores principales: | , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
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The American Society for Cell Biology
2016
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5170604/ https://www.ncbi.nlm.nih.gov/pubmed/27708139 http://dx.doi.org/10.1091/mbc.E16-04-0224 |
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author | Génier, Samuel Degrandmaison, Jade Moreau, Pierrick Labrecque, Pascale Hébert, Terence E. Parent, Jean-Luc |
author_facet | Génier, Samuel Degrandmaison, Jade Moreau, Pierrick Labrecque, Pascale Hébert, Terence E. Parent, Jean-Luc |
author_sort | Génier, Samuel |
collection | PubMed |
description | Mechanisms that prevent aggregation and promote folding of nascent G protein–coupled receptors (GPCRs) remain poorly understood. We identified chaperonin containing TCP-1 subunit eta (CCT7) as an interacting partner of the β-isoform of thromboxane A(2) receptor (TPβ) by yeast two-hybrid screening. CCT7 coimmunoprecipitated with overexpressed TPβ and β(2)-adrenergic receptor (β(2)AR) in HEK 293 cells, but also with endogenous β(2)AR. CCT7 depletion by small interfering RNA reduced total and cell-surface expression of both receptors and caused redistribution of the receptors to juxtanuclear aggresomes, significantly more so for TPβ than β(2)AR. Interestingly, Hsp90 coimmunoprecipitated with β(2)AR but virtually not with TPβ, indicating that nascent GPCRs can adopt alternative folding pathways. In vitro pull-down assays showed that both receptors can interact directly with CCT7 through their third intracellular loops and C-termini. We demonstrate that Trp(334) in the TPβ C-terminus is critical for the CCT7 interaction and plays an important role in TPβ maturation and cell-surface expression. Of note, introducing a tryptophan in the corresponding position of the TPα isoform confers the CCT7-binding and maturation properties of TPβ. We show that an interaction with a subunit of the CCT/TCP-1 ring complex (TRiC) chaperonin complex is involved in regulating aggregation of nascent GPCRs and in promoting their proper maturation and expression. |
format | Online Article Text |
id | pubmed-5170604 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2016 |
publisher | The American Society for Cell Biology |
record_format | MEDLINE/PubMed |
spelling | pubmed-51706042017-02-16 Regulation of GPCR expression through an interaction with CCT7, a subunit of the CCT/TRiC complex Génier, Samuel Degrandmaison, Jade Moreau, Pierrick Labrecque, Pascale Hébert, Terence E. Parent, Jean-Luc Mol Biol Cell Articles Mechanisms that prevent aggregation and promote folding of nascent G protein–coupled receptors (GPCRs) remain poorly understood. We identified chaperonin containing TCP-1 subunit eta (CCT7) as an interacting partner of the β-isoform of thromboxane A(2) receptor (TPβ) by yeast two-hybrid screening. CCT7 coimmunoprecipitated with overexpressed TPβ and β(2)-adrenergic receptor (β(2)AR) in HEK 293 cells, but also with endogenous β(2)AR. CCT7 depletion by small interfering RNA reduced total and cell-surface expression of both receptors and caused redistribution of the receptors to juxtanuclear aggresomes, significantly more so for TPβ than β(2)AR. Interestingly, Hsp90 coimmunoprecipitated with β(2)AR but virtually not with TPβ, indicating that nascent GPCRs can adopt alternative folding pathways. In vitro pull-down assays showed that both receptors can interact directly with CCT7 through their third intracellular loops and C-termini. We demonstrate that Trp(334) in the TPβ C-terminus is critical for the CCT7 interaction and plays an important role in TPβ maturation and cell-surface expression. Of note, introducing a tryptophan in the corresponding position of the TPα isoform confers the CCT7-binding and maturation properties of TPβ. We show that an interaction with a subunit of the CCT/TCP-1 ring complex (TRiC) chaperonin complex is involved in regulating aggregation of nascent GPCRs and in promoting their proper maturation and expression. The American Society for Cell Biology 2016-12-01 /pmc/articles/PMC5170604/ /pubmed/27708139 http://dx.doi.org/10.1091/mbc.E16-04-0224 Text en © 2016 Génier et al. This article is distributed by The American Society for Cell Biology under license from the author(s). Two months after publication it is available to the public under an Attribution–Noncommercial–Share Alike 3.0 Unported Creative Commons License (http://creativecommons.org/licenses/by-nc-sa/3.0). “ASCB®,” “The American Society for Cell Biology®,” and “Molecular Biology of the Cell®” are registered trademarks of The American Society for Cell Biology. |
spellingShingle | Articles Génier, Samuel Degrandmaison, Jade Moreau, Pierrick Labrecque, Pascale Hébert, Terence E. Parent, Jean-Luc Regulation of GPCR expression through an interaction with CCT7, a subunit of the CCT/TRiC complex |
title | Regulation of GPCR expression through an interaction with CCT7, a subunit of the CCT/TRiC complex |
title_full | Regulation of GPCR expression through an interaction with CCT7, a subunit of the CCT/TRiC complex |
title_fullStr | Regulation of GPCR expression through an interaction with CCT7, a subunit of the CCT/TRiC complex |
title_full_unstemmed | Regulation of GPCR expression through an interaction with CCT7, a subunit of the CCT/TRiC complex |
title_short | Regulation of GPCR expression through an interaction with CCT7, a subunit of the CCT/TRiC complex |
title_sort | regulation of gpcr expression through an interaction with cct7, a subunit of the cct/tric complex |
topic | Articles |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5170604/ https://www.ncbi.nlm.nih.gov/pubmed/27708139 http://dx.doi.org/10.1091/mbc.E16-04-0224 |
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