The α-Arrestin ARRDC3 Regulates the Endosomal Residence Time and Intracellular Signaling of the β2-Adrenergic Receptor

Tian, Xufan; Irannejad, Roshanak; Bowman, Shanna L.; Du, Yang; Puthenveedu, Manojkumar A.; Zastrow, Mark von; Benovic, Jeffrey L.

doi:10.1074/jbc.m116.716589

Cited by 60 publications

(57 citation statements)

References 43 publications

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“…3B). The ␤ 2 AR displayed comparable colocation with LAMP1 after prolonged agonist stimulation (33). These studies suggest that activated and internalized PAR4 sorts to early endosomes and then to late endosomes/lysosomes.…”

Section: Internalized Par4 Is Trafficked Through Early Endosomesmentioning

confidence: 55%

Protease-activated Receptor-4 Signaling and Trafficking Is Regulated by the Clathrin Adaptor Protein Complex-2 Independent of β-Arrestins

Smith

Coronel

et al. 2016

Journal of Biological Chemistry

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Protease-activated receptor-4 (PAR4) is a G protein-coupled receptor (GPCR) for thrombin and is proteolytically activated, similar to the prototypical PAR1. Due to the irreversible activation of PAR1, receptor trafficking is intimately linked to signal regulation. However, unlike PAR1, the mechanisms that control PAR4 trafficking are not known. Here, we sought to define the mechanisms that control PAR4 trafficking and signaling. In HeLa cells depleted of clathrin by siRNA, activated PAR4 failed to internalize. Consistent with clathrin-mediated endocytosis, expression of a dynamin dominant-negative K44A mutant also blocked activated PAR4 internalization. However, unlike most GPCRs, PAR4 internalization occurred independently of ␤-arrestins and the receptor's C-tail domain. Rather, we discovered a highly conserved tyrosine-based motif in the third intracellular loop of PAR4 and found that the clathrin adaptor protein complex-2 (AP-2) is important for internalization. Depletion of AP-2 inhibited PAR4 internalization induced by agonist. In addition, mutation of the critical residues of the tyrosine-based motif disrupted agonist-induced PAR4 internalization. Using Dami megakaryocytic cells, we confirmed that AP-2 is required for agonist-induced internalization of endogenous PAR4. Moreover, inhibition of activated PAR4 internalization enhanced ERK1/2 signaling, whereas Akt signaling was markedly diminished. These findings indicate that activated PAR4 internalization requires AP-2 and a tyrosine-based motif and occurs independent of ␤-arrestins, unlike most classical GPCRs. Moreover, these findings are the first to show that internalization of activated PAR4 is linked to proper ERK1/2 and Akt activation.Thrombin is a coagulant protease that plays an essential role in hemostasis and thrombosis in response to tissue injury (1).Thrombin is locally generated at sites of vascular injury, where it initiates responses in various cell types including platelets, endothelial, and smooth muscle cells. Thrombin elicits cellular responses through members of the protease-activated receptor (PAR) 5 family of G protein-coupled receptors (GPCRs). Unlike classic GPCRs, which are activated by a diffusible ligand, PARs are activated by the proteolytic cleavage of the receptor N-terminal domain (2, 3). The newly exposed N terminus, termed the "tethered ligand," then binds intramolecularly to the receptor's second extracellular loop, inducing a conformational change that facilitates coupling to heterotrimeric G proteins and downstream signaling. Studies have shown that a six-amino acid synthetic peptide mimic of a PAR's tethered ligand is sufficient to activate the receptor and evoke cellular responses similar to those resulting from proteolytic cleavage of the receptor (4). Because of the irreversible mechanism of PAR1 activation with the generation of a tethered ligand that cannot diffuse away, the mechanisms that regulate PAR1 trafficking are important for maintaining proper signaling and appropriate cellular responses (5).After activa...

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Section: Internalized Par4 Is Trafficked Through Early Endosomesmentioning

confidence: 55%

Protease-activated Receptor-4 Signaling and Trafficking Is Regulated by the Clathrin Adaptor Protein Complex-2 Independent of β-Arrestins

Smith

Coronel

et al. 2016

Journal of Biological Chemistry

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“…How PPP1R14C interacts with Vps35 and modulates endosomal PTHR signaling remains poorly understood but may relate to delayed entry into ASRT endosomal tubules, resulting in prolonged endosomal residence time, enhanced intracellular PTHR signaling and eventual misdirection into degradative lysosomes as observed in this study [65]. In fact, this mechanism has been recently shown to underscore the ability of arrestin domain-containing protein 3 (ARRDC3), a member of the β-arrestin family of visual/β-arrestins and Vps26-related proteins [66] to prolong on the endosomal occupancy and associated cAMP-signaling of β 2 AR [67]. Although future work will be required to appreciate the full complement of proteins that may intersect with the ASRT-signaling-trafficking nexus, it appears that in addition to its canonical function in directing entry of receptors into recycling tubules, the ASRT complex, together with accessory proteins, is capable of integrating endosomal GPCR-signal termination with post-endocytic sorting.…”

Section: Asrt-mediated Endosome-to-plasma Membrane Recyclingmentioning

confidence: 67%

GPCR Signaling and Trafficking: The Long and Short of It

Pavlos

Friedman

2017

Trends in Endocrinology & Metabolism

170

121

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Emerging findings disclose unexpected components of G protein-coupled receptor (GPCR) signaling and cell biology. Select GPCRs exhibit classical signaling that is restricted to cell membranes and newly described persistent signaling that depends on internalization of the GPCR bound to β-arrestins. Termination of non-canonical endosomal signaling requires intraluminal acidification and sophisticated protein trafficking machineries. Recent studies reveal the structural determinants of the trafficking chaperones. This review summarizes advances in GPCR signaling and trafficking with a focus on the parathyroid hormone receptor as prototype, and the actin-SNX27-retromer tubule complex, an endosomal sorting hub responsible for recycling and preservation of cell surface receptors. The findings are integrated into a model of PTHR trafficking with implications for signal transduction, bone growth, and mineral-ion metabolism.

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“…ARRDC3 has been shown to regulate endosomal sorting of the b2AR, a classic GPCR (36)(37)(38). However, the precise role of ARRDC3 in this pathway remains controversial given the dominant role of b-arrestins in regulation of b2AR function (36,38).…”

Section: Discussionmentioning

confidence: 99%

“…The a-arrestin ARRDC3 shares structural homology to the b-arrestin family of adaptor proteins (47), which have important regulatory roles in mammalian GPCR signaling and trafficking (37). ARRDC3 has been shown to regulate endosomal sorting of the b2AR, a classic GPCR (36)(37)(38).…”

Section: Discussionmentioning

confidence: 99%

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The α-arrestin ARRDC3 suppresses breast carcinoma invasion by regulating G protein–coupled receptor lysosomal sorting and signaling

Arakaki

Pan

Lin

et al. 2018

Journal of Biological Chemistry

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Aberrant G protein-coupled receptor (GPCR) expression and activation has been linked to tumor initiation, progression, invasion and metastasis. However, compared with other cancer drivers, the exploitation of GPCRs as potential therapeutic targets has been largely ignored, despite the fact that GPCRs are highly druggable. Therefore, to advance the potential status of GPCRs as therapeutic targets, it is important to understand how GPCRs function together with other cancer drivers during tumor progression. We now report that the a-arrestin domain-containing protein-3 (ARRDC3) acts as a tumor suppressor in part by controlling signaling and trafficking of the GPCR, protease-activated receptor-1 (PAR1). In a series of highly invasive basal-like breast carcinomas, we found that expression of ARRDC3 is suppressed while PAR1 is aberrantly overexpressed because of defective lysosomal sorting that results in persistent signaling. Using a lentiviral doxycycline-inducible system, we demonstrate that re-expression of ARRDC3 in invasive breast carcinoma is sufficient to restore normal PAR1 trafficking through the ALGinteracting protein X (ALIX)-dependent lysosomal degradative pathway. We also show that ARRDC3 re-expression attenuates PAR1-stimulated persistent signaling of c-Jun NH2-terminal kinase (JNK) in invasive breast cancer. Remarkably, restoration of ARRDC3 expression significantly reduced activated PAR1-induced breast carcinoma invasion, which was also dependent on JNK signaling. These findings are the first to identify a critical link between the tumor suppressor ARRDC3 and regulation of GPCR trafficking and signaling in breast cancer.G protein-coupled receptors (GPCRs) are a large family of cell surface signaling receptors that play a critical role in cancer growth and development by regulating cellular proliferation, invasion, migration, immune cell-mediated functions, angiogenesis and survival at metastatic sites (1-3). GPCR function can be altered in cancer through aberrant overexpression, gain-of-function activating mutations, mutations in downstream G protein signaling effectors, and increased production and secretion of GPCR activating ligands by both tumor cells and surrounding stromal cells (4-7). As cell surface receptors with highly druggable sites, GPCRs are the largest class of drug targets, with over 30% of current FDAhttp://www.jbc.org/cgi

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The α-Arrestin ARRDC3 Regulates the Endosomal Residence Time and Intracellular Signaling of the β2-Adrenergic Receptor

Cited by 60 publications

References 43 publications

Protease-activated Receptor-4 Signaling and Trafficking Is Regulated by the Clathrin Adaptor Protein Complex-2 Independent of β-Arrestins

Protease-activated Receptor-4 Signaling and Trafficking Is Regulated by the Clathrin Adaptor Protein Complex-2 Independent of β-Arrestins

GPCR Signaling and Trafficking: The Long and Short of It

The α-arrestin ARRDC3 suppresses breast carcinoma invasion by regulating G protein–coupled receptor lysosomal sorting and signaling

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