Reconstituting regulation of the canonical Wnt pathway by engineering a minimal β-catenin destruction machine

Pronobis, Mira I.; Deuitch, Natalie; Posham, Vinya; Mimori-Kiyosue, Yuko; Peifer, Mark

doi:10.1091/mbc.e16-07-0557

Cited by 26 publications

(41 citation statements)

References 38 publications

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“…These findings support a model in which multiple direct and indirect interactions between components redundantly cooperate to enhance robustness of the destruction complex. These redundancy features of the axin complex critically depend on interactions with APC (Pronobis et al, ).…”

Section: Axin Is the Primary Coordinator Of Destruction Complex Activitymentioning

confidence: 99%

“…This region of APC harbours a number of short axin and β‐catenin binding motifs as well as regulatory regions essential for β‐catenin proteolysis, as discussed below. At its very C‐terminus, APC carries a basic domain that promotes actin assembly (Okada et al, ) and a microtubule interaction region, both of which are dispensable for β‐catenin degradation (Smits et al, ; McCartney and Nathke, ; Pronobis et al, ). Overall, regulatory interactions of APC with the cytoskeleton are thought to mediate alternative roles of APC in spindle formation, kinetochore attachment, microtubule stability as well as the regulation of cell motility and polarity (Nathke, ; Okada et al, ).…”

Section: Essential Role Of Apc In the Destruction Complexmentioning

confidence: 99%

“…Due to the presence of multiple binding sites for a single partner as well as overlap in self‐oligomerization capacity, axin and APC appear to partially share redundant functions inside the destruction complex. To identify the essential parts of both scaffold proteins, a recent study compiled a minimal destruction complex by using only five essential regions of axin and APC (Pronobis et al, ). For APC, these regions included the self‐associating ASAD domain, Arm repeats and the 20R2‐CID region.…”

Section: The Axin–apc Interaction Is Highly Dynamicmentioning

confidence: 99%

“…These APC regions were coupled to the axin C‐terminus containing the β‐catenin binding domain and DIX domain. The artificial scaffold protein formed cytosolic puncta and allowed full restoration of β‐catenin destruction in APC‐mutant SW480 cells (Pronobis et al, ). While these results are highly informative, it should be noted that these experiments relied on overexpression and were performed in the presence of endogenous wild‐type axin as well as truncated APC, both of which could contribute to the formation and activity of the destruction complex.…”

Section: The Axin–apc Interaction Is Highly Dynamicmentioning

confidence: 99%

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Molecular regulation and pharmacological targeting of the β‐catenin destruction complex

Kappel

Maurice

2017

British J Pharmacology

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The β‐catenin destruction complex is a dynamic cytosolic multiprotein assembly that provides a key node in Wnt signalling regulation. The core components of the destruction complex comprise the scaffold proteins axin and adenomatous polyposis coli and the Ser/Thr kinases casein kinase 1 and glycogen synthase kinase 3. In unstimulated cells, the destruction complex efficiently drives degradation of the transcriptional coactivator β‐catenin, thereby preventing the activation of the Wnt/β‐catenin pathway. Mutational inactivation of the destruction complex is a major pathway in the pathogenesis of cancer. Here, we review recent insights in the regulation of the β‐catenin destruction complex, including newly identified interaction interfaces, regulatory elements and post‐translationally controlled mechanisms. In addition, we discuss how mutations in core destruction complex components deregulate Wnt signalling via distinct mechanisms and how these findings open up potential therapeutic approaches to restore destruction complex activity in cancer cells.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

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Section: Axin Is the Primary Coordinator Of Destruction Complex Activitymentioning

confidence: 99%

Section: Essential Role Of Apc In the Destruction Complexmentioning

confidence: 99%

Section: The Axin–apc Interaction Is Highly Dynamicmentioning

confidence: 99%

Section: The Axin–apc Interaction Is Highly Dynamicmentioning

confidence: 99%

See 2 more Smart Citations

Molecular regulation and pharmacological targeting of the β‐catenin destruction complex

Kappel

Maurice

2017

British J Pharmacology

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“…Condensate formation is driven by Axin polymerization via its DIX domain, by APC function, and by other multivalent interactions e.g. (Fagotto et al, 1999;Kishida et al, 1999;Cliffe et al, 2003;Schwarz-Romond et al, 2007a;Faux et al, 2008;Mendoza-Topaz et al, 2011;Kunttas-Tatli et al, 2014;Thorvaldsen et al, 2015) (Pronobis et al, 2015;Pronobis et al, 2017).…”

Section: Introductionmentioning

confidence: 99%

Wnt Regulation: Exploring Axin-Disheveled interactions and defining mechanisms by which the SCF E3 ubiquitin ligase is recruited to the destruction complex

Schaefer

Pronobis

Williams

et al. 2019

Preprint

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Wnt signaling plays key roles in embryonic development and adult stem cell homeostasis, and is altered in human cancer. Signaling is turned on and off by regulating stability of the effector β-catenin. The multiprotein destruction complex binds and phosphorylates β-catenin, and transfers it to the SCF-TrCP E3-ubiquitin ligase, for ubiquitination and destruction. Wnt signals act though Dishevelled to turn down the destruction complex, stabilizing β-catenin. Recent work clarified underlying mechanisms, but important questions remain. We explore β-catenin transfer from the destruction complex to the E3 ligase, and test models suggesting Dishevelled and APC2 compete for association with Axin. We find that Slimb/TrCP is a dynamic component of the destruction complex biomolecular condensate, while other E3 proteins are not. Recruitment requires Axin and not APC, and Axin's RGS domain plays an important role. We find that elevating Dishevelled levels in Drosophila embryos has paradoxical effects, promoting the ability of limiting levels of Axin to turn off Wnt signaling. When we elevate Dishevelled levels, it forms its own cytoplasmic puncta, but these do not recruit Axin. SIM imaging in mammalian cells suggests that this may result by promoting Dishevelled: Dishevelled interactions at the expense of Dishevelled:Axin interactions when Dishevelled levels are high.part of the destruction complex, either by direct interaction with destruction complex proteins or using phosphorylated βcat as a bridge. In this model, once βcat is phosphorylated it could be directly transferred to the E3 ligase, thus preventing dephosphorylation of βcat by cellular phosphatases during transit. Immunoprecipitation (IP) experiments in animals and cell culture revealed that βTrCP can coIP with Axin, APC, βcat, and GSK3, and that Wnt signals reduce Axin: βTrCP coIP (Hart et al., 1999;Kitagawa et al., 1999;Liu et al., 1999;Li et al., 2012). However, these studies did not examine whether βTrCP or other components of the E3 are recruited to the destruction complex, leaving both models an option, especially if βTrCP acts as a shuttling protein between complexes. Here we address this issue.A second set of outstanding questions concern the mechanisms by which Wnt signaling down-regulates βcat destruction. Wnt signaling is initiated when Wnt ligands interact with complex multi-protein receptors, comprised of Frizzled family members plus LRP5/6 (reviewed in (DeBruine et al., 2017;Nusse and Clevers, 2017). This receptor complex recruits the destruction complex to the plasma membrane, via interaction of Axin with the phosphorylated LRP5/6 tail and with the Wnt effector Disheveled (Dvl in mammals/Dsh in Drosophila).This leads to downregulation of the destruction complex, reducing the rate of βcat destruction. Current data suggest destruction complex downregulation occurs via multiple mechanisms (reviewed in (MacDonald and He, 2012;Nusse and Clevers, 2017), some rapid and others initiated more slowly. These include direct inhibition of GSK3 by the phosphor...

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Liquid–liquid phase separation drives the β‐catenin destruction complex formation

2021

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The intracellular multiprotein complex β-catenin destruction complex plays a key role in Wnt/β-catenin signaling. Wnt stimulation induces the assembly of the receptorassociated signalosome and the inactivation of the destruction complex, leading to β-catenin accumulation and transcriptional activation of the target genes. The core components of the destruction complex include Axin, APC, GSK3β, CK1α and other proteins. Recent studies demonstrated that Axin and APC undergo liquid-liquid phase separation (LLPS), which is critical for their function to regulate Wnt/β-catenin signaling. Here, we discuss the possible roles of LLPS in Wnt/β-catenin signaling and regulation of Axin LLPS by post-translational modifications.

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Reconstituting regulation of the canonical Wnt pathway by engineering a minimal β-catenin destruction machine

Cited by 26 publications

References 38 publications

Molecular regulation and pharmacological targeting of the β‐catenin destruction complex

Molecular regulation and pharmacological targeting of the β‐catenin destruction complex

Wnt Regulation: Exploring Axin-Disheveled interactions and defining mechanisms by which the SCF E3 ubiquitin ligase is recruited to the destruction complex

Liquid–liquid phase separation drives the β‐catenin destruction complex formation

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