Essential role of the Dishevelled DEP domain in a Wnt-dependent human-cell-based complementation assay

Gammons, Melissa V.; Rutherford, Trevor J.; Steinhart, Zachary; Angers, Stéphane; Bienz, Mariann

doi:10.1242/jcs.195685

Cited by 72 publications

(116 citation statements)

References 65 publications

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“…We tested the effects of the Dvl2 DIX structure-based mutants in a luciferasebased reporter (TOPFLASH) assay that measures Wnt-stimulated b-catenin/TCFstimulated transcription (Molenaar et al, 1996), expressing mutant Dvl2 constructs in Dvl triple knockout (TKO) cells (Gammons et al, 2016b) cells at near-endogenous levels and normalizing the luciferase signal by the expression level of the mutant relative to wild-type Dvl2 (Fig. 3c, S2).…”

Section: Forms An Antiparallel Double Helix That Stabilizes Filammentioning

confidence: 99%

“…3c, 5), indicating that oligomerization of Dvl is necessary for signal transduction. However, the extent to which Dvl oligomerizes at endogenous levels has not been established (Gammons et al, 2016b;Ma et al, 2020;Schaefer et al, 2018;Smalley et al, 2005). Using HEK293T cells in which a C-terminal GFP fusion was knocked into the endogenous Dvl2 locus, we performed live epifluorescent imaging with and without Wnt3A stimulation.…”

Section: The Size Of Dvl Oligomers Is Limited In Cellsmentioning

confidence: 99%

“…The Dvl-Axin interaction is essential for Wnt/b-catenin signal transduction and is mediated by a DIX (Dvl and Axin interacting) domain present in each protein (Cliffe et al, 2003;Fiedler et al, 2011;Julius et al, 2000;Kishida et al, 1999;Smalley et al, 1999). Dvl has an N-terminal DIX domain, as well as a PDZ and a DEP domain; the DEP domain mediates the interaction with Fzd (Gammons et al, 2016b;Tauriello et al, 2012). Axin contains binding sites for APC, GSK-3, b-catenin and CK1 (Behrens et al, 1998;Hart et al, 1998;Ikeda et al, 1998;Kishida et al, 1999) and a C-terminal DIX domain.…”

Section: Introductionmentioning

confidence: 99%

“…The extent of filament/oligomer formation by endogenous Dvl and Axin, and how their relative stoichiometries in signaling complexes control GSK-3 activity and b-catenin destruction, are essential but unresolved questions (Gammons et al, 2016b;Schaefer et al, 2018). Here, we examine the interaction of Axin and Dvl DIX domains, using structural and biochemical analyses of purified DIX and DAX domains, and assessing the role of these domains when the proteins are present at endogenous levels.…”

Section: Introductionmentioning

confidence: 99%

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Limited Dishevelled/Axin oligomerization determines efficiency of Wnt/β-catenin signal transduction

Kan

Enos

Korkmazhan

et al. 2020

Preprint

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SUMMARYIn Wnt/β-catenin signaling, the transcriptional coactivator β-catenin is regulated by its phosphorylation in a complex that includes the scaffold protein Axin and associated kinases. Wnt binding to its coreceptors activates the cytosolic effector Dishevelled (Dvl), leading to the recruitment of Axin and the inhibition of β-catenin phosphorylation. This process requires interaction of homologous DIX domains present in Dvl and Axin, but is mechanistically undefined. We show that Dvl DIX forms antiparallel, double-stranded oligomers in vitro, and that Dvl in cells forms oligomers typically <10 molecules at endogenous expression levels. Axin DIX (DAX) forms small single-stranded oligomers, but its self-association is stronger than that of DIX. DAX caps the ends of DIX oligomers, such that a DIX oligomer has at most four DAX binding sites. The relative affinities and stoichiometry of the DIX-DAX interaction provide a mechanism for efficient inhibition of β-catenin phosphorylation upon Axin recruitment to the Wnt receptor complex.

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Section: Forms An Antiparallel Double Helix That Stabilizes Filammentioning

confidence: 99%

Section: The Size Of Dvl Oligomers Is Limited In Cellsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Limited Dishevelled/Axin oligomerization determines efficiency of Wnt/β-catenin signal transduction

Kan

Enos

Korkmazhan

et al. 2020

Preprint

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“…The DEP domain binds the intracellular core of the FZD TMD upon receptor activation and undergoes a highly thermostable domain swap (formation of an intermolecular β-sheet) to form dimers (Gammons et al, 2016a). It is possible that coordination of DEP domains at FZD dimers is a requirement for this domain-swapping event (Gammons et al, 2016b). We also note that domain-swapped DEP dimers Wnt signalosome assembly BJP British Journal of Pharmacology (2017) 174 4564-4574 4571 formed tetrameric assemblies in two different crystal forms (Gammons et al, 2016a).…”

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confidence: 99%

Assembly and architecture of the Wnt/β‐catenin signalosome at the membrane

DeBruine

Melcher

2017

British J Pharmacology

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Wnt/β-catenin signalling is initiated by a ternary Wnt-Frizzled (FZD)-LDL receptor-related protein (LRP) 5/6 binding event. The resulting conformational changes in the FZD and LRP5/6 receptors promote the assembly of an intracellular signalosome driven by Dishevelled and Axin co-polymerization. Recent evidence suggests that the FZD receptor and LRP5/6 participate in the assembly of this signalosome by forming regulatory scaffolds for stabilizing Dishevelled and Axin adapters. In this review, we focus on the contributions of Wnts and their receptors in the assembly of the signalosome. We present an emerging model, which unifies Wnt receptor oligomerization with intracellular signalosome formation, and then discuss how FZD receptors might be targeted to either disrupt or enhance their capacity as a dynamic sensor of Wnt binding. LINKED ARTICLESThis article is part of a themed section on WNT Signalling: Mechanisms and Therapeutic Opportunities. To view the other articles in this section visit http://onlinelibrary.wiley.com/doi/10.1111/bph.v174.24/issuetoc IntroductionWingless/int1 (Wnt) signal activation is tightly controlled by a dynamic signalosome consisting of Class Frizzled GPCRs (FZDs), LDL receptor-related protein (LRP) 5/6 coreceptors and Dishevelled and Axin adapters (Cong et al., 2004;Kaykas et al., 2004;Bilic et al., 2007;Gammons et al., 2016a). In this review, we discuss emerging evidence for a model of FZD and LRP5/6 co-oligomerization. In this model, oligomerization of the Wnt signalling complex is facilitated by domains with weak homo-oligomerization propensities such as the FZD cysteine-rich domain (CRD), the LRP5/6 β-propeller (βP) domains, and the Dishevelled EGL-10 and pleckstrin (DEP) and Dishevelled and Axin (DIX) domains (Dann et al., 2001;Gammons et al., 2016b). In the absence of Wnt, inactive complexes of FZD, LRP5/6 and DVL pose as static sensors for Wnt binding (Chen et al., 2014) ( Figure 1A). When Wnt binds FZD receptors and LRP5/6, the FZD receptor and LRP5/6 complexes are activated and oligomerize to create an extensive scaffold for Dishevelled stabilization and interaction with LRP5/6 (Bilic et al., 2007) ( Figure 1B). Dishevelled then co-polymerizes with Axin through shared DIX domains to form a trap which sequesters the β-catenin destruction complex (Schwarz-Romond et al., 2007;Fiedler et al., 2011).Consequentially, β-catenin translocates to the nucleus where it works in concert with Wnt transcription factors to turn on Wnt target genes (van de Wetering et al., 1991). Wnt family ligands are FZD-specific, highly conserved and often determine developmental patterning and cell fate (van de Wetering et al., 1997). Many of these physiological consequences are the result of Wnt/β-catenin signalling which involves the assembly of an intracellular Dishevelled/Axin signalosome (Cong et al., 2004; SchwarzRomond et al., 2007). Unlike norrin, an atypical FZD 4 / LRP5 agonist, all 19 human Wnts share a highly conserved two-domain structure which enables it to attach the FZD receptor CRD and bin...

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Wnt signalosomes: What we know that we do not know

Hartmann,

Siddiqui,

Bryant

et al. 2024

BioEssays

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Signaling through the Wnt/β‐catenin pathway is relayed through three multiprotein complexes: (1) the membrane‐associated signalosome, which includes the activated Wnt receptors, (2) the cytoplasmic destruction complex that regulates turnover of the transcriptional coactivator β‐catenin, and (3) the nuclear enhanceosome that mediates pathway‐specific transcription. Recent discoveries have revealed that Wnt receptor activities are tightly regulated to maintain proper tissue homeostasis and that aberrant receptor upregulation enhances Wnt signaling to drive tumorigenesis, highlighting the importance of signalosome control. These studies have focused on the detailed process by which Wnt ligands engage their coreceptors, LRP5/6 and Frizzled. However, the components that constitute the signalosome and the regulation of their assembly remain undefined. In this review, we discuss Wnt/β‐catenin signalosome composition and the mechanisms that regulate signalosome assembly, including the role of biomolecular condensates and ubiquitylation. We also summarize the evidence for the presence of Wnt ligand‐independent signalosome formation.

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Essential role of the Dishevelled DEP domain in a Wnt-dependent human-cell-based complementation assay

Cited by 72 publications

References 65 publications

Limited Dishevelled/Axin oligomerization determines efficiency of Wnt/β-catenin signal transduction

Limited Dishevelled/Axin oligomerization determines efficiency of Wnt/β-catenin signal transduction

Assembly and architecture of the Wnt/β‐catenin signalosome at the membrane

Wnt signalosomes: What we know that we do not know

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