Noncanonical interactions of G proteins and β‐arrestins: from competitors to companions

Smith, Jeffrey S.; Pack, Thomas F.

doi:10.1111/febs.15749

Cited by 12 publications

(3 citation statements)

References 115 publications

(128 reference statements)

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“…For several GPCRs, β‐arrestin 1 and 2 proteins are required for ERK activation 32–34 and previously we also showed that NF‐α1/CPE activated 5‐HTR1E can increase ERK phosphorylation via β‐arrestin recruitment 4 . Here we investigated if serotonin‐5‐HTR1E mediated ERK phosphorylation is also β‐arrestin dependent.…”

Section: Resultsmentioning

confidence: 79%

Characterization of serotonin‐5‐HTR1E signaling pathways and its role in cell survival

et al. 2023

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5-Hydroxytryptamine receptor 1E(5-HTR1E) is reported to activate cyclic AMP (cAMP) and extracellular-signal related kinases (ERK) pathways via its ligands and binding partners, but the detailed mechanism underlying the serotonininduced 5-HTR1E signaling is still not known. In the present study, we determined the cellular regulators of ERK and cAMP signaling pathways in response to serotonin-induced 5-HTR1E activation in 5-HTR1E overexpressing HEK293 cells.We found that Pertussis Toxin (PTX) treatment completely reversed the effect of serotonin-5-HTR1E mediated signaling on cAMP and ERK pathways, confirming the involvement of a Gαi-linked cascade. We also observed that Gβγ and Gq were not associated with 5-HTR1E activation, while blocking protein kinase A (PKA) inhibited ERK signaling only, and had no effect on cAMP. Additionally, serotoninstimulated ERK1/2 phosphorylation was similar in 5-HTR1E overexpressing, βarrestin-deficient HEK293 cells and is solely dependent on G protein signaling. siRNA mediated gene knockdown studies in SH-SY5Y cells revealed that the inhibition of 5-HTR1E reduced the expression of cMyc, Cyclin D1, Cyclin E and BCL2 genes which are related to cell cycle regulation and survival. MTT assays showed that 5-HTR1E knockdown in SHSY-5Y and U118 cells inhibited cell survival significantly. In addition to the signaling mechanism, we also performed RNA-seq analysis in 5-HTR1E overexpressing HEK293 cells and found that 5-HTR1E can regulate the expression of Receptor activity modifying protein 1 (RAMP1), Nuclear receptor 1 (NR4A1) and other Cyclin genes. These findings indicate that serotonin interaction with 5-HTR1E receptor simultaneously activates cAMP and ERK pathway in HEK293 cells and its expression is important for cell survival.

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Section: Resultsmentioning

confidence: 79%

Characterization of serotonin‐5‐HTR1E signaling pathways and its role in cell survival

et al. 2023

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“…For several GPCRs, β-arrestin 1 and 2 proteins are required for the activation of ERK pathway (31)(32)(33) and previously we also showed that NF-α1/CPE activated 5-HTR1E can increase ERK phosphorylation via β-arrestin recruitment ( 4). Here we explored if serotonin-5-HTR1E mediated ERK phosphorylation is also β-arrestin dependent.…”

Section: Serotonin-5-htr1e Activated Erk Pathway Is β-Arrestin Indepe...mentioning

confidence: 74%

Characterization of serotonin-5-HTR1E signaling pathways and its role in cell survival

Sharma¹,

Campbell²,

Yang³

et al. 2023

Preprint

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5-Hydroxy tryptamine receptor 1E (5-HTR1E) is reported to activate cAMP and ERK pathways via its ligands and binding partners, but the detailed mechanism underlying the serotonin induced 5-HTR1E signaling is still not known. In the present study, we determined the cellular regulators of ERK and cAMP signaling pathways in response to serotonin induced 5-HTR1E activation in 5-HTR1E overexpressing HEK293 cells. We found that Pertussis Toxin (PTX) treatment completely reversed the effect of serotonin-5-HTR1E mediated signaling on cAMP and ERK pathways, confirming the involvement of a Gαi-linked cascade. We also observed that Gβγ and Gq were not associated with 5-HTR1E activation, while blocking PKA inhibited ERK signaling only, and had no effect on cAMP. Additionally, serotonin-stimulated ERK1/2 phosphorylation was similar in 5-HTR1E overexpressing, β-arrestin-deficient HEK293 cells and is solely dependent on G protein signaling. siRNA mediated gene knockout studies in SH-SY5Y cells revealed that the inhibition of 5-HTR1E reduced the expression of cMyc, Cyclin D1, Cyclin E and BCL2 genes which are related to cell cycle regulation and survival. MTT assays showed that 5-HTR1E knockdown in SHSY-5Y and U118 cells inhibited cell survival significantly. In addition to the signaling mechanism, we also performed RNA-seq analysis in 5-HTR1E overexpressing HEK293 cells and found that 5-HTR1E can regulate the expression of Receptor activity modifying protein 1 (RAMP1), Nuclear receptor 1 (NR4A1) and other Cyclin genes. These findings indicate that serotonin interaction with 5-HTR1E receptor simultaneously activates cAMP and ERK pathway in HEK293 cells and its expression is important for cell survival.

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“…Noncanonical GPCR signaling is an emerging area of study that adds to our understanding of the complexity of GPCR signaling and how a single agonist:receptor interaction results in the activation of complex cellular and physiologic processes. For instance, recent findings suggest that Gαi can form a complex with β-arrestin and scaffold MAPK, even at receptors that do not canonically couple to Gαi ( 23, 70 ). There have also been reported ternary complexes between G proteins, β-arrestins, and GPCRs that can modulate GPCR signaling ( 22, 71, 72 ).…”

Section: Discussionmentioning

confidence: 99%

ACKR3 Proximity Labeling Identifies Novel G protein- and β-arrestin-independent GPCR Interacting Proteins

Hicks,

Gardner,

Eiger

et al. 2024

Preprint

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The canonical paradigm of GPCR signaling recognizes G proteins and β-arrestins as the two primary transducers that promote GPCR signaling. Recent evidence suggests the atypical chemokine receptor 3 (ACKR3) does not couple to G proteins, and β-arrestins are dispensable for some of its functions. Here, we employed proximity labeling to identify proteins that interact with ACKR3 in cells devoid of β-arrestin. We identified proteins involved in the endocytic machinery and evaluated a subset of proteins conserved across several GPCR-based proximity labeling experiments. We discovered that the bone morphogenic protein 2-inducible kinase (BMP2K) interacts with many different GPCRs with varying dependency on β-arrestin. Together, our work highlights the existence of modulators that can act independently of G proteins and β-arrestins to regulate GPCR signaling and provides important evidence for other targets that may regulate GPCR signaling.

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Noncanonical interactions of G proteins and β‐arrestins: from competitors to companions

Cited by 12 publications

References 115 publications

Characterization of serotonin‐5‐HTR1E signaling pathways and its role in cell survival

Characterization of serotonin‐5‐HTR1E signaling pathways and its role in cell survival

Characterization of serotonin-5-HTR1E signaling pathways and its role in cell survival

ACKR3 Proximity Labeling Identifies Novel G protein- and β-arrestin-independent GPCR Interacting Proteins

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