β-arrestin-dependent, G protein-independent ERK1/2 activation by the β2 adrenergic receptor

Sudha K. Shenoy, Matthew M Drake, Christopher D. Nelson, Daniel A. Houtz, Kunhong Xiao, Srinivasan Madabushi, Eric Reiter, Richard T. Premont, Olivier Lichtarge, Robert J. Lefkowitz

Research output: Contribution to journalArticle

496 Citations (Scopus)

Abstract

Physiological effects of β adrenergic receptor (β2AR) stimulation have been classically shown to result from G s-dependent adenylyl cyclase activation. Here we demonstrate a novel signaling mechanism wherein β-arrestins mediate β2AR signaling to extracellular-signal regulated kinases 1/2 (ERK 1/2) independent of G protein activation. Activation of ERK1/2 by the β2AR expressed in HEK-293 cells was resolved into two components dependent, respectively, on G s-G i/protein kinase A (PKA) or β-arrestins. G protein-dependent activity was rapid, peaking within 2-5 min, was quite transient, was blocked by pertussis toxin (G i inhibitor) and H-89 (PKA inhibitor), and was insensitive to depletion of endogenous β-arrestins by siRNA. β-Arrestin-dependent activation was slower in onset (peak 5-10 min), less robust, but more sustained and showed little decrement over 30 min. It was insensitive to pertussis toxin and H-89 and sensitive to depletion of either β-arrestin1 or -2 by small interfering RNA. In G s knock-out mouse embryonic fibroblasts, wild-type β2AR recruited β-arrestin2-green fluorescent protein and activated pertussis toxin-insensitive ERK1/2. Furthermore, a novel β2AR mutant (β2AR T68F,Y132G,Y219A or β2AR TYY), rationally designed based on Evolutionary Trace analysis, was incapable of G protein activation but could recruit β-arrestins, undergo β-arrestin-dependent internalization, and activate β-arrestin-dependent ERK. Interestingly, overexpression of GRK5 or -6 increased mutant receptor phosphorylation and β-arrestin recruitment, led to the formation of stable receptor-β- arrestin complexes on endosomes, and increased agonist-stimulated phospho-ERK1/2. In contrast, GRK2, membrane translocation of which requires Gβγ release upon G protein activation, was ineffective unless it was constitutively targeted to the plasma membrane by a prenylation signal (CAAX). These findings demonstrate that the β2AR can signal to ERK via a GRK5/6-β-arrestin-dependent pathway, which is independent of G protein coupling.

Original languageEnglish (US)
Pages (from-to)1261-1273
Number of pages13
JournalJournal of Biological Chemistry
Volume281
Issue number2
DOIs
StatePublished - Jan 13 2006
Externally publishedYes

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Arrestin
Arrestins
GTP-Binding Proteins
Adrenergic Receptors
Chemical activation
Pertussis Toxin
Cyclic AMP-Dependent Protein Kinases
Small Interfering RNA
Prenylation
Trace analysis
Phosphorylation
Mitogen-Activated Protein Kinase 3
HEK293 Cells
Endosomes
Mitogen-Activated Protein Kinase 1
Fibroblasts
Cell membranes
Protein Kinase Inhibitors
Green Fluorescent Proteins
Adenylyl Cyclases

ASJC Scopus subject areas

  • Biochemistry

Cite this

Shenoy, S. K., Drake, M. M., Nelson, C. D., Houtz, D. A., Xiao, K., Madabushi, S., ... Lefkowitz, R. J. (2006). β-arrestin-dependent, G protein-independent ERK1/2 activation by the β2 adrenergic receptor. Journal of Biological Chemistry, 281(2), 1261-1273. https://doi.org/10.1074/jbc.M506576200

β-arrestin-dependent, G protein-independent ERK1/2 activation by the β2 adrenergic receptor. / Shenoy, Sudha K.; Drake, Matthew M; Nelson, Christopher D.; Houtz, Daniel A.; Xiao, Kunhong; Madabushi, Srinivasan; Reiter, Eric; Premont, Richard T.; Lichtarge, Olivier; Lefkowitz, Robert J.

In: Journal of Biological Chemistry, Vol. 281, No. 2, 13.01.2006, p. 1261-1273.

Research output: Contribution to journalArticle

Shenoy, SK, Drake, MM, Nelson, CD, Houtz, DA, Xiao, K, Madabushi, S, Reiter, E, Premont, RT, Lichtarge, O & Lefkowitz, RJ 2006, 'β-arrestin-dependent, G protein-independent ERK1/2 activation by the β2 adrenergic receptor', Journal of Biological Chemistry, vol. 281, no. 2, pp. 1261-1273. https://doi.org/10.1074/jbc.M506576200
Shenoy, Sudha K. ; Drake, Matthew M ; Nelson, Christopher D. ; Houtz, Daniel A. ; Xiao, Kunhong ; Madabushi, Srinivasan ; Reiter, Eric ; Premont, Richard T. ; Lichtarge, Olivier ; Lefkowitz, Robert J. / β-arrestin-dependent, G protein-independent ERK1/2 activation by the β2 adrenergic receptor. In: Journal of Biological Chemistry. 2006 ; Vol. 281, No. 2. pp. 1261-1273.
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