Chapter One - Ubiquitination and Deubiquitination of G Protein-Coupled Receptors

Jean-Charles, Pierre-Yves; Snyder, Joshua C.; Shenoy, Sudha K.

doi:10.1016/bs.pmbts.2016.05.001

Cited by 34 publications

(31 citation statements)

References 239 publications

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“…Regardless of whether or not Fzds are true GPCRs, this does not preclude them from functioning in a manner similar to GPCRs. GPCR desensitization is regulated by internalization of the receptor, a process that is dependent on posttranslational modifications such as phosphorylation (Jean-Charles et al, 2016). In our study, we have found that Fzd9b is phosphorylated on tyrosine residues in response to Wnt9a, consistent with a role for the receptor tyrosine kinase EGFR in this process.…”

Section: A Model For Wnt-fzd Signaling Specificitysupporting

confidence: 74%

EGFR confers exquisite specificity of Wnt9a-Fzd9b signaling in hematopoietic stem cell development

Grainger

Nguyen

Richter

et al. 2018

Preprint

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SummaryThe mechanisms of Wnt-Frizzled (Fzd) signaling selectivity and their biological implications remain unclear. We demonstrate for the first time that the epidermal growth factor receptor (EGFR) is required as a co-factor for Wnt signaling. Using genetic studies in zebrafish, paired with in vitro cell biology and biochemistry, we have determined that Fzd9b signals specifically with Wnt9a in vivo and in vitro to elicit β-catenin dependent Wnt signals that regulate hematopoietic stem and progenitor cell (HSPC) development in the dorsal aorta. This requirement is conserved in the derivation of HSPCs from human embryonic stem cells. Wnt9a-Fzd9b specificity requires two intracellular domains in Fzd9b, which interact with EGFR as a required co-factor to promote signal transduction. EGFR phosphorylates one tyrosine residue on Fzd9b, a requirement for the Wnt signal. These findings indicate that Wnt signaling interactions can be exquisitely specific and inform protocols for derivation of HSPCs in vitro.HighlightsAn in vitro signaling screen identifies Fzd9b as a Wnt9a-specific receptor.Fzd9b and Wnt9a regulate hematopoietic stem cell development as a cognate pair.WNT9A and FZD9 are required for HSPC derivation from human pluripotent cells in vitro.EGFR confers specificity to Wnt9a-Fzd9b signaling in zebrafish and human cells.

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Section: A Model For Wnt-fzd Signaling Specificitysupporting

confidence: 74%

EGFR confers exquisite specificity of Wnt9a-Fzd9b signaling in hematopoietic stem cell development

Grainger

Nguyen

Richter

et al. 2018

Preprint

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“…2C). For a number of GPCRs, ubiquitination serves as a sorting signal to promote their trafficking to late endosomes and lysosomes (19,25). We therefore tested whether USP20-mediated deubiquitination of the ␤ 1 AR affects lysosomal trafficking of internalized ␤ 1 ARs.…”

Section: Usp20 Functions As a Cognate Deubiquitinase For The ␤ 1 Armentioning

confidence: 99%

“…GPCR lysosomal trafficking and degradation can also be modulated by ubiquitination, a post-translational modification that appends monomers or polymeric chains of the 76-amino acid protein ubiquitin to substrate proteins (18,19). Although the dynamics and mechanisms of ␤ 2 AR ubiquitination have been broadly characterized (20 -24), the mechanisms that regulate ␤ 1 AR ubiquitination and associated vesicular trafficking are largely unknown.…”

mentioning

confidence: 99%

The deubiquitinase ubiquitin–specific protease 20 is a positive modulator of myocardial β1-adrenergic receptor expression and signaling

Yu¹,

Jean-Charles²,

Abraham

et al. 2019

Journal of Biological Chemistry

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Reversible ubiquitination of G protein-coupled receptors regulates their trafficking and signaling; whether deubiquitinases regulate myocardial ␤ 1 -adrenergic receptors (␤ 1 ARs) is unknown.Wereportthatubiquitin-specificprotease20(USP20)deubiquitinates and attenuates lysosomal trafficking of the ␤ 1 AR. ␤ 1 AR-induced phosphorylation of USP20 Ser-333 by protein kinase A-␣ (PKA␣) was required for optimal USP20-mediated regulation of ␤ 1 AR lysosomal trafficking. Both phosphomimetic (S333D) and phosphorylation-impaired (S333A) USP20 possess intrinsic deubiquitinase activity equivalent to WT activity. However, unlike USP20 WT and S333D, the S333A mutant associated poorly with the ␤ 1 AR and failed to deubiquitinate the ␤ 1 AR. USP20 -KO mice showed normal baseline systolic function but impaired ␤ 1 AR-induced contractility and relaxation. Dobutamine stimulation did not increase cAMP in USP20 -KO left ventricles (LVs), whereas NKH477-induced adenylyl cyclase activity was equivalent to WT. The USP20 homolog USP33, which shares redundant roles with USP20, had no effect on ␤ 1 AR ubiquitination, but USP33 was up-regulated in USP20 -KO hearts suggesting compensatory regulation. Myocardial ␤ 1 AR expression in USP20 -KO was drastically reduced, whereas ␤ 2 AR expression was maintained as determined by radioligand binding in LV sarcolemmal membranes. Phospho-USP20 was significantly increased in LVs of wildtype (WT) mice after a 1-week catecholamine infusion and a 2-week chronic pressure overload induced by transverse aortic constriction (TAC). Phospho-USP20 was undetectable in ␤ 1 AR KO mice subjected to TAC, suggesting a role for USP20 phosphorylation in cardiac response to pressure overload. We conclude that USP20 regulates ␤ 1 AR signaling in vitro and in vivo. Additionally, ␤ 1 AR-induced USP20 phosphorylation may serve as a feedforward mechanism to stabilize ␤ 1 AR expression and signaling during pathological insults to the myocardium. by guest on July 10, 2020 http://www.jbc.org/ Downloaded from Figure 1. Agonist-induced ubiquitination and lysosomal trafficking of the ␤ 1 AR.A, HEK-293 cells stably transfected with FLAG-␤ 1 AR were stimulated with 1 M isoproterenol (Iso) for indicated times and subjected to FLAG IP followed by serial immunoblotting (IB) with anti-ubiquitin antibody (rabbit polyclonal, Bethyl Laboratories Inc.) and polyclonal FLAG antibody. The 1st lane shows the background signal obtained from HEK-293 cells transfected with vector. B, ubiquitin smears were quantitated and normalized to cognate FLAG-␤ 1 AR bands and plotted as % maximum signal (see "Experimental procedures"). The graph includes means Ϯ S.E. from four independent experiments. *, p Ͻ 0.05 compared with 0 min, one-way ANOVA, and Bonferroni's test. C, HEK-293 cells with stable transfection of ␤ 1 AR-CFP were stimulated with 1 M Iso for the indicated times, and the distribution of ␤ 1 AR (red) and LAMP1 (green) was visualized with LSM-510 confocal microscope. Representative images are shown, and quantification of colocalization from Ն13 z...

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“…Of the 600 E3 ligases, the majority are classified as RING E3 ligases, whereas the HECT E3 ligases are a considerably smaller class and includes 28 members. The NEDD4 (neural precursor cell expressed, developmentally downregulated‐4) HECT domain‐containing E3 ligases subfamily is comprised of nine members and are best known to mediate ubiquitination and endo‐lysosomal sorting of GPCRs following ligand stimulation . PAR2 is an exception and ubiquitinated by c‐Cbl, a RING E3 ligase, in an agonist‐dependent manner and required for endo‐lysosomal sorting .…”

Section: Ubiquitin E3 Ligases Function In Direct and Indirect Regulatmentioning

confidence: 99%

“…PAR2 is an exception and ubiquitinated by c‐Cbl, a RING E3 ligase, in an agonist‐dependent manner and required for endo‐lysosomal sorting . In contrast to NEDD4 E3 ligase, the RING E3 ligases have been largely reported to mediate constitutive ubiquitination of several GPCRs and appear to regulate a diverse array of trafficking processes . Despite the large number of studies describing ligand‐induced GPCR ubiquitination, our knowledge of the mechanisms by which GPCRs regulate E3 ligase activity to facilitate receptor endo‐lysosomal sorting is limited.…”

Section: Ubiquitin E3 Ligases Function In Direct and Indirect Regulatmentioning

confidence: 99%

Endo‐lysosomal sorting of G‐protein‐coupled receptors by ubiquitin: Diverse pathways for G‐protein‐coupled receptor destruction and beyond

Dores

Trejo

2018

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Ubiquitin is covalently attached to substrate proteins in the form of a single ubiquitin moiety or polyubiquitin chains and has been generally linked to protein degradation, however, distinct types of ubiquitin linkages are also used to control other critical cellular processes like cell signaling. Over forty mammalian G protein‐coupled receptors (GPCRs) have been reported to be ubiquitinated, but despite the diverse and rich complexity of GPCR signaling, ubiquitin has been largely ascribed to receptor degradation. Indeed, GPCR ubiquitination targets the receptors for degradation by lysosome, which is mediated by the Endosomal Sorting Complexes Required for Transport (ESCRT) machinery, and the proteasome. This has led to the view that ubiquitin and ESCRTs primarily function as the signal to target GPCRs for destruction. Contrary to this conventional view, studies indicate that ubiquitination of certain GPCRs and canonical ubiquitin‐binding ESCRTs are not required for receptor degradation and revealed that diverse and complex pathways exist to regulate endo‐lysosomal sorting of GPCRs. In other studies, GPCR ubiquitination has been shown to drive signaling and not receptor degradation and further revealed novel insight into the mechanisms by which GPCRs trigger the activity of the ubiquitination machinery. Here, we discuss the diverse pathways by which ubiquitin controls GPCR endo‐lysosomal sorting and beyond.

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Chapter One - Ubiquitination and Deubiquitination of G Protein-Coupled Receptors

Cited by 34 publications

References 239 publications

EGFR confers exquisite specificity of Wnt9a-Fzd9b signaling in hematopoietic stem cell development

EGFR confers exquisite specificity of Wnt9a-Fzd9b signaling in hematopoietic stem cell development

The deubiquitinase ubiquitin–specific protease 20 is a positive modulator of myocardial β1-adrenergic receptor expression and signaling

Endo‐lysosomal sorting of G‐protein‐coupled receptors by ubiquitin: Diverse pathways for G‐protein‐coupled receptor destruction and beyond

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