Seeing β-arrestin in action: The role of β-arrestins in Histamine 1 Receptor signaling

Pietraszewska-Bogiel, Anna; Goedhart, Joachim

doi:10.1101/775049

Cited by 4 publications

(4 citation statements)

References 60 publications

(70 reference statements)

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“…H1R DRY-mediated EKAR signals showed, however, a much faster decline than the H1R-mediated signals. Treatment with a specific G q protein inhibitor, YM254890, reduced H1R-mediated EKARcyt signals (Figure B), in agreement with the Gq-dependent activation of the ERK pathway downstream of H1R . Interestingly, inhibition with YM254890 completely abolished the EKARcyt signal in cells expressing H1R DRY receptors (Figure B), suggesting that the ERK signal induced by the DRY mutant requires the activation of Gq.…”

Section: Resultssupporting

confidence: 65%

Not So Dry After All: DRY Mutants of the AT1_A Receptor and H1 Receptor Can Induce G-Protein-Dependent Signaling

et al. 2020

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G-protein-coupled receptors (GPCRs) are seven transmembrane spanning receptors that regulate a wide array of intracellular signaling cascades in response to various stimuli. To do so, they couple to different heterotrimeric G proteins and adaptor proteins, including arrestins. Importantly, arrestins were shown to regulate GPCR signaling through G proteins, as well as promote G protein-independent signaling events. Several research groups have reported successful isolation of exclusively G protein-dependent and arrestin-dependent signaling downstream of GPCR activation using biased agonists or receptor mutants incapable of coupling to either arrestins or G proteins. In the latter category, the DRY mutant of the angiotensin II type 1 receptor was extensively used to characterize the functional selectivity downstream of AT1AR. In an attempt to understand histamine 1 receptor signaling, we characterized the signaling capacity of the H1R DRY mutant in a panel of dynamic, live cell biosensor assays, including arrestin recruitment, heterotrimeric G protein activation, Ca2+ signaling, protein kinase C activity, GTP binding of RhoA, and activation of ERK1/2. Here, we show that both H1R DRY mutant and the AT1AR DRY mutant are capable of efficient activation of G protein-mediated signaling. Therefore, contrary to the common belief, they do not constitute suitable tools for the dissection of the arrestin-mediated, G protein-independent signaling downstream of these receptors.

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Section: Resultssupporting

confidence: 65%

Not So Dry After All: DRY Mutants of the AT1_A Receptor and H1 Receptor Can Induce G-Protein-Dependent Signaling

et al. 2020

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“…We selected three agonists that would activate three different families of GPCRs that are endogenously present in HeLa cells. Histamine is reported to predominantly activate Gq in HeLa cells by the histamine H1 receptor ( Pietraszewska-Bogiel and Goedhart, 2019 preprint), and UK activates Gi by α2-adrenergic receptors ( van Unen et al, 2016b ). Our data with the inhibitors YM and PTx, which are selective for Gq and Gi, respectively, show that the two agonists indeed preferentially activate a single heterotrimeric G-protein class.…”

Section: Discussionmentioning

confidence: 99%

Single-cell imaging of ERK and Akt activation dynamics and heterogeneity induced by G-protein-coupled receptors

Chavez-Abiega

Grönloh

Gadella

et al. 2022

Journal of Cell Science

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Kinases play key roles in signaling networks that are activated by G protein-coupled receptors (GPCRs). Kinase activities are generally inferred from cell lysates, hiding cell-to-cell variability. To study the dynamics and heterogeneity of ERK and Akt, we employed high-content biosensor imaging with kinase translocation reporters. The kinases were activated with GPCR ligands. We observed ligand-concentration dependent response kinetics to histamine, α2-adrenergic, and S1P receptor stimulation. By using G protein inhibitors, we observed that Gq mediated the ERK and Akt responses to histamine. In contrast, Gi was necessary for ERK and Akt activation in response to α2-adrenergic receptor activation. ERK and Akt were also strongly activated by S1P, showing high heterogeneity at the single cell level, especially for ERK. Cluster analysis of time-series derived from 68,000 cells obtained under the different conditions revealed several distinct populations of cells that display similar response dynamics. ERK response dynamics to S1P showed high heterogeneity, which was reduced by the inhibition of Gi. To conclude, we have set up an imaging and analysis strategy that reveals substantial cell-to-cell heterogeneity in kinase activity driven by GPCRs.

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“…We selected three agonists that would activate three different families of GPCRs that are endogenously present in HeLa cells. Histamine is reported to predominantly activate Gq in HeLa cells by the histamine H1 receptor [36] and UK activates Gi by α2-adrenergic receptors [37]. Our data with the inhibitors YM and PTx, which are selective of Gq and Gi respectively, show that the two agonists indeed preferentially activate a single heterotrimeric G protein class.…”

Section: Discussionmentioning

confidence: 55%

Heterogeneity and dynamics of ERK and Akt activation by G protein-coupled receptors depend on the activated heterotrimeric G proteins

Chavez-Abiega

Grönloh

Gadella

et al. 2021

Preprint

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Kinases are fundamental regulators of cellular functions and play key roles in GPCR-mediated signaling pathways. Kinase activities are generally inferred from cell lysates, hiding the heterogeneity of the individual cellular responses to extracellular stimuli. Here, we study the dynamics and heterogeneity of ERK and Akt in genetically identical cells in response to activation of endogenously expressed GPCRs. We use kinase translocation reporters, high-content imaging, automated segmentation and clustering methods to assess cell-to-cell signaling heterogeneity. We observed ligand-concentration dependent response kinetics to histamine, α2-adrenergic, and S1P receptor stimulation that varied between cells. By using G protein inhibitors, we observed that Gq mediated the ERK and Akt responses to histamine. In contrast, Gi was necessary for ERK and Akt activation in response to α2-adrenergic receptor activation. ERK and Akt were also strongly activated by S1P, showing high heterogeneity at the single cell level, especially for ERK. In all cases, the cellular heterogeneity was not explained by distinct pre-stimulation levels or saturation of the measured response. Cluster analysis of time-series derived from 68,000 cells obtained under the different conditions revealed several distinct populations of cells that display similar response dynamics. The single-cell ERK responses to histamine and UK showed remarkably similar dynamics, despite the activation of different heterotrimeric G proteins. In contrast, the ERK response dynamics to S1P showed high heterogeneity, which was reduced by the inhibition of Gi. To conclude, we have set up an imaging and analysis strategy that reveals substantial cell-to-cell heterogeneity in kinase activity driven by GPCRs.

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Seeing β-arrestin in action: The role of β-arrestins in Histamine 1 Receptor signaling

Cited by 4 publications

References 60 publications

Not So Dry After All: DRY Mutants of the AT1_A Receptor and H1 Receptor Can Induce G-Protein-Dependent Signaling

Not So Dry After All: DRY Mutants of the AT1_A Receptor and H1 Receptor Can Induce G-Protein-Dependent Signaling

Single-cell imaging of ERK and Akt activation dynamics and heterogeneity induced by G-protein-coupled receptors

Heterogeneity and dynamics of ERK and Akt activation by G protein-coupled receptors depend on the activated heterotrimeric G proteins

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Seeing β-arrestin in action: The role of β-arrestins in Histamine 1 Receptor signaling

Cited by 4 publications

References 60 publications

Not So Dry After All: DRY Mutants of the AT1A Receptor and H1 Receptor Can Induce G-Protein-Dependent Signaling

Not So Dry After All: DRY Mutants of the AT1A Receptor and H1 Receptor Can Induce G-Protein-Dependent Signaling

Single-cell imaging of ERK and Akt activation dynamics and heterogeneity induced by G-protein-coupled receptors

Heterogeneity and dynamics of ERK and Akt activation by G protein-coupled receptors depend on the activated heterotrimeric G proteins

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Not So Dry After All: DRY Mutants of the AT1_A Receptor and H1 Receptor Can Induce G-Protein-Dependent Signaling

Not So Dry After All: DRY Mutants of the AT1_A Receptor and H1 Receptor Can Induce G-Protein-Dependent Signaling