Beta(2)-adrenoceptors increase translocation of GLUT4 via GPCR kinase sites in the receptor C-terminal tail

Nodi Dehvari, Dana Hutchinson, Julia Nevzorova, Olof Dallner, Masaaki Sato, Martina Kocan, Jon Merlin, Bronwyn Evans, Roger Summers, Tore Bengtsson

Research output: Contribution to journalArticleResearchpeer-review

Abstract

Background and purpose: beta-Adrenoceptor (beta-AR) stimulation induces glucose uptake in several insulin-sensitive tissues, by poorly understood mechanisms. Experimental approach: We used a model system in CHO-K1 (Chinese Hamster ovary K1) cells expressing the human beta(2) -AR and glucose transporter 4 (GLUT4) to investigate the signalling mechanisms involved. Key results: In CHO-K1 cells there was no response to beta-AR agonists. The introduction of beta(2) -ARs and GLUT4 into these cells caused increased glucose uptake in response to beta-AR agonists. GLUT4 translocation occurred in response to insulin and beta(2) -AR stimulation, although the key insulin signalling intermediate Akt was not phosphorylated in response to beta(2) -AR stimulation. Truncation of the C-terminus of the beta(2) -AR at position 349 to remove known phosphorylation sites for G protein-coupled receptor kinases (GRKs) or at position 344 to remove an additional protein kinase A (PKA) site together with the GRK phosphorylation sites did not significantly affect cyclic AMP accumulation but decreased beta(2) -AR stimulated glucose uptake. Furthermore, inhibition of GRK by transfection of betaARKct inhibited beta(2) -AR-mediated glucose uptake and GLUT4 translocation, and over-expression of a kinase-dead GRK2 mutant (GRK2 K220R) also inhibited GLUT4 translocation. Introducing beta(2) -AR lacking phosphorylation sites for GRK or PKA demonstrated that the GRK sites, but not the PKA sites, were necessary for GLUT4 translocation. Conclusions and implications: Glucose uptake in response to activation of beta(2) -ARs involves translocation of GLUT4 in this model system. The mechanism is dependent on the C-terminus of the beta(2) -AR, requires GRK phosphorylation sites, and involves a signalling pathway distinct from that stimulated by insulin.
Original languageEnglish
Pages (from-to)1442 - 1456
Number of pages15
JournalBritish Journal of Pharmacology
Volume165
Issue number5
DOIs
Publication statusPublished - 2012

Cite this

Dehvari, Nodi ; Hutchinson, Dana ; Nevzorova, Julia ; Dallner, Olof ; Sato, Masaaki ; Kocan, Martina ; Merlin, Jon ; Evans, Bronwyn ; Summers, Roger ; Bengtsson, Tore. / Beta(2)-adrenoceptors increase translocation of GLUT4 via GPCR kinase sites in the receptor C-terminal tail. In: British Journal of Pharmacology. 2012 ; Vol. 165, No. 5. pp. 1442 - 1456.
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title = "Beta(2)-adrenoceptors increase translocation of GLUT4 via GPCR kinase sites in the receptor C-terminal tail",
abstract = "Background and purpose: beta-Adrenoceptor (beta-AR) stimulation induces glucose uptake in several insulin-sensitive tissues, by poorly understood mechanisms. Experimental approach: We used a model system in CHO-K1 (Chinese Hamster ovary K1) cells expressing the human beta(2) -AR and glucose transporter 4 (GLUT4) to investigate the signalling mechanisms involved. Key results: In CHO-K1 cells there was no response to beta-AR agonists. The introduction of beta(2) -ARs and GLUT4 into these cells caused increased glucose uptake in response to beta-AR agonists. GLUT4 translocation occurred in response to insulin and beta(2) -AR stimulation, although the key insulin signalling intermediate Akt was not phosphorylated in response to beta(2) -AR stimulation. Truncation of the C-terminus of the beta(2) -AR at position 349 to remove known phosphorylation sites for G protein-coupled receptor kinases (GRKs) or at position 344 to remove an additional protein kinase A (PKA) site together with the GRK phosphorylation sites did not significantly affect cyclic AMP accumulation but decreased beta(2) -AR stimulated glucose uptake. Furthermore, inhibition of GRK by transfection of betaARKct inhibited beta(2) -AR-mediated glucose uptake and GLUT4 translocation, and over-expression of a kinase-dead GRK2 mutant (GRK2 K220R) also inhibited GLUT4 translocation. Introducing beta(2) -AR lacking phosphorylation sites for GRK or PKA demonstrated that the GRK sites, but not the PKA sites, were necessary for GLUT4 translocation. Conclusions and implications: Glucose uptake in response to activation of beta(2) -ARs involves translocation of GLUT4 in this model system. The mechanism is dependent on the C-terminus of the beta(2) -AR, requires GRK phosphorylation sites, and involves a signalling pathway distinct from that stimulated by insulin.",
author = "Nodi Dehvari and Dana Hutchinson and Julia Nevzorova and Olof Dallner and Masaaki Sato and Martina Kocan and Jon Merlin and Bronwyn Evans and Roger Summers and Tore Bengtsson",
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Beta(2)-adrenoceptors increase translocation of GLUT4 via GPCR kinase sites in the receptor C-terminal tail. / Dehvari, Nodi; Hutchinson, Dana; Nevzorova, Julia; Dallner, Olof; Sato, Masaaki; Kocan, Martina; Merlin, Jon; Evans, Bronwyn; Summers, Roger; Bengtsson, Tore.

In: British Journal of Pharmacology, Vol. 165, No. 5, 2012, p. 1442 - 1456.

Research output: Contribution to journalArticleResearchpeer-review

TY - JOUR

T1 - Beta(2)-adrenoceptors increase translocation of GLUT4 via GPCR kinase sites in the receptor C-terminal tail

AU - Dehvari, Nodi

AU - Hutchinson, Dana

AU - Nevzorova, Julia

AU - Dallner, Olof

AU - Sato, Masaaki

AU - Kocan, Martina

AU - Merlin, Jon

AU - Evans, Bronwyn

AU - Summers, Roger

AU - Bengtsson, Tore

PY - 2012

Y1 - 2012

N2 - Background and purpose: beta-Adrenoceptor (beta-AR) stimulation induces glucose uptake in several insulin-sensitive tissues, by poorly understood mechanisms. Experimental approach: We used a model system in CHO-K1 (Chinese Hamster ovary K1) cells expressing the human beta(2) -AR and glucose transporter 4 (GLUT4) to investigate the signalling mechanisms involved. Key results: In CHO-K1 cells there was no response to beta-AR agonists. The introduction of beta(2) -ARs and GLUT4 into these cells caused increased glucose uptake in response to beta-AR agonists. GLUT4 translocation occurred in response to insulin and beta(2) -AR stimulation, although the key insulin signalling intermediate Akt was not phosphorylated in response to beta(2) -AR stimulation. Truncation of the C-terminus of the beta(2) -AR at position 349 to remove known phosphorylation sites for G protein-coupled receptor kinases (GRKs) or at position 344 to remove an additional protein kinase A (PKA) site together with the GRK phosphorylation sites did not significantly affect cyclic AMP accumulation but decreased beta(2) -AR stimulated glucose uptake. Furthermore, inhibition of GRK by transfection of betaARKct inhibited beta(2) -AR-mediated glucose uptake and GLUT4 translocation, and over-expression of a kinase-dead GRK2 mutant (GRK2 K220R) also inhibited GLUT4 translocation. Introducing beta(2) -AR lacking phosphorylation sites for GRK or PKA demonstrated that the GRK sites, but not the PKA sites, were necessary for GLUT4 translocation. Conclusions and implications: Glucose uptake in response to activation of beta(2) -ARs involves translocation of GLUT4 in this model system. The mechanism is dependent on the C-terminus of the beta(2) -AR, requires GRK phosphorylation sites, and involves a signalling pathway distinct from that stimulated by insulin.

AB - Background and purpose: beta-Adrenoceptor (beta-AR) stimulation induces glucose uptake in several insulin-sensitive tissues, by poorly understood mechanisms. Experimental approach: We used a model system in CHO-K1 (Chinese Hamster ovary K1) cells expressing the human beta(2) -AR and glucose transporter 4 (GLUT4) to investigate the signalling mechanisms involved. Key results: In CHO-K1 cells there was no response to beta-AR agonists. The introduction of beta(2) -ARs and GLUT4 into these cells caused increased glucose uptake in response to beta-AR agonists. GLUT4 translocation occurred in response to insulin and beta(2) -AR stimulation, although the key insulin signalling intermediate Akt was not phosphorylated in response to beta(2) -AR stimulation. Truncation of the C-terminus of the beta(2) -AR at position 349 to remove known phosphorylation sites for G protein-coupled receptor kinases (GRKs) or at position 344 to remove an additional protein kinase A (PKA) site together with the GRK phosphorylation sites did not significantly affect cyclic AMP accumulation but decreased beta(2) -AR stimulated glucose uptake. Furthermore, inhibition of GRK by transfection of betaARKct inhibited beta(2) -AR-mediated glucose uptake and GLUT4 translocation, and over-expression of a kinase-dead GRK2 mutant (GRK2 K220R) also inhibited GLUT4 translocation. Introducing beta(2) -AR lacking phosphorylation sites for GRK or PKA demonstrated that the GRK sites, but not the PKA sites, were necessary for GLUT4 translocation. Conclusions and implications: Glucose uptake in response to activation of beta(2) -ARs involves translocation of GLUT4 in this model system. The mechanism is dependent on the C-terminus of the beta(2) -AR, requires GRK phosphorylation sites, and involves a signalling pathway distinct from that stimulated by insulin.

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U2 - 10.1111/j.1476-5381.2011.01647.x

DO - 10.1111/j.1476-5381.2011.01647.x

M3 - Article

VL - 165

SP - 1442

EP - 1456

JO - British Journal of Pharmacology

JF - British Journal of Pharmacology

SN - 1476-5381

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