Lack of beta-arrestin signaling in the absence of active G proteins.
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Abstract |
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G protein-independent, arrestin-dependent signaling is a paradigm that broadens the signaling scope of G protein-coupled receptors (GPCRs) beyond G proteins for numerous biological processes. However, arrestin signaling in the collective absence of functional G proteins has never been demonstrated. Here we achieve a state of "zero functional G" at the cellular level using HEK293 cells depleted by CRISPR/Cas9 technology of the Gs/q/12 families of Gα proteins, along with pertussis toxin-mediated inactivation of Gi/o. Together with HEK293 cells lacking β-arrestins ("zero arrestin"), we systematically dissect G protein- from arrestin-driven signaling outcomes for a broad set of GPCRs. We use biochemical, biophysical, label-free whole-cell biosensing and ERK phosphorylation to identify four salient features for all receptors at "zero functional G": arrestin recruitment and internalization, but-unexpectedly-complete failure to activate ERK and whole-cell responses. These findings change our understanding of how GPCRs function and in particular of how they activate ERK1/2. |
Year of Publication |
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2018
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Journal |
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Nature communications
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Volume |
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9
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Issue |
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1
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Number of Pages |
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341
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Date Published |
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2018
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URL |
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http://dx.doi.org/10.1038/s41467-017-02661-3
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DOI |
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10.1038/s41467-017-02661-3
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Short Title |
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Nat Commun
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