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

Schaefer, Kristina N.; Pronobis, Mira I.; Williams, Clara E.; Zhang, Shiping; Bauer, Lauren; Goldfarb, Dennis; Yan, Feng; Major, Michael B.; Peifer, Mark

doi:10.1091/mbc.e19-11-0647

Cited by 18 publications

(7 citation statements)

References 80 publications

(209 reference statements)

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“…AXIN1, however, appears to interact with SCF β-TrCP much more weakly than APC, at least under our experimental conditions. These observations differ from those of Schaefer et al. (2020) , in which the Drosophila β-TrCP ortholog Slimb was shown to co-localize and co-immunoprecipitate with Axin but not Apc2.…”

Section: Discussioncontrasting

confidence: 95%

“…Omission of β-TrCP abolished the elution shift by APC ( Figure 6 ), suggesting that the interaction either occurs directly through β-TrCP or through another β-TrCP-dependent mechanism. A recently proposed interaction of Drosophila Axin with the β-TrCP ortholog Slimb ( Schaefer et al., 2020 ) prompted us to perform the same experiment with AXIN1. However, we observed only a small population of SCF β-TrCP shifting compared with APC ( Figure S8 D), indicating that APC is the predominant DC component binding to the E3 complex in our system.…”

Section: Resultsmentioning

confidence: 99%

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Reconstitution of the destruction complex defines roles of AXIN polymers and APC in β-catenin capture, phosphorylation, and ubiquitylation

et al. 2021

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Summary The Wnt/β-catenin pathway is a highly conserved, frequently mutated developmental and cancer pathway. Its output is defined mainly by β-catenin’s phosphorylation- and ubiquitylation-dependent proteasomal degradation, initiated by the multi-protein β-catenin destruction complex. The precise mechanisms underlying destruction complex function have remained unknown, largely because of the lack of suitable in vitro systems. Here we describe the in vitro reconstitution of an active human β-catenin destruction complex from purified components, recapitulating complex assembly, β-catenin modification, and degradation. We reveal that AXIN1 polymerization and APC promote β-catenin capture, phosphorylation, and ubiquitylation. APC facilitates β-catenin’s flux through the complex by limiting ubiquitylation processivity and directly interacts with the SCF β-TrCP E3 ligase complex in a β-TrCP-dependent manner. Oncogenic APC truncation variants, although part of the complex, are functionally impaired. Nonetheless, even the most severely truncated APC variant promotes β-catenin recruitment. These findings exemplify the power of biochemical reconstitution to interrogate the molecular mechanisms of Wnt/β-catenin signaling.

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Section: Discussioncontrasting

confidence: 95%

Section: Resultsmentioning

confidence: 99%

Reconstitution of the destruction complex defines roles of AXIN polymers and APC in β-catenin capture, phosphorylation, and ubiquitylation

et al. 2021

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“…The biocondensate model of β-catenin destruction complex has been proposed recently 12,33 . The biocondensates create physically separated compartments in the cytosol that is specifically enriched of the components of the complex.…”

Section: Discussionmentioning

confidence: 99%

Multivalent tumor suppressor adenomatous polyposis coli promotes Axin biomolecular condensate formation and efficient β-catenin degradation

Ren

Husmann

et al. 2020

Sci Rep

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The tumor suppressor adenomatous polyposis coli (APC) is frequently mutated in colorectal cancers. APC and Axin are core components of a destruction complex that scaffolds GSK3β and CK1 to earmark β-catenin for proteosomal degradation. Disruption of APC results in pathologic stabilization of β-catenin and oncogenesis. However, the molecular mechanism by which APC promotes β-catenin degradation is unclear. Here, we find that the intrinsically disordered region (IDR) of APC, which contains multiple β-catenin and Axin interacting sites, undergoes liquid–liquid phase separation (LLPS) in vitro. Expression of the APC IDR in colorectal cells promotes Axin puncta formation and β-catenin degradation. Our results support the model that multivalent interactions between APC and Axin drives the β-catenin destruction complex to form biomolecular condensates in cells, which concentrate key components to achieve high efficient degradation of β-catenin.

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“…The phosphorylation leads to the ubiquitination and degradation of β-catenin. If Wnt binds to the transmembrane complex, the protein Disheveled is activated and turns down the destruction complex, resulting in accumulation of β-catenin in the cytoplasm[ 9 , 10 ]. Then, β-catenin is translocated into the nucleus and acts there as a transcription factor together with P300, B-cell CLL/lymphoma 9, pygo and T-cell factor/lymphoid enhancer-binding factor (TCF/LEF) as cofactors[ 11 - 13 ].…”

Section: Wnt/β-catenin Signaling Pathwaymentioning

confidence: 99%

Intestinal Wnt in the transition from physiology to oncology

Swoboda¹,

Mittelsdorf²,

Chen³

et al. 2022

WJCO

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Adult stem cells are necessary for self-renewal tissues and regeneration after damage. Especially in the intestine, which self-renews every few days, they play a key role in tissue homeostasis. Therefore, complex regulatory mechanisms are needed to prevent hyperproliferation, which can lead in the worst case to carcinogenesis or under-activation of stem cells, which can result in dysfunctional epithelial. One main regulatory signaling pathway is the Wnt/β-catenin signaling pathway. It is a highly conserved pathway, with β-catenin, a transcription factor, as target protein. Translocation of β-catenin from cytoplasm to nucleus activates the transcription of numerous genes involved in regulating stem cell pluripo-tency, proliferation, cell differentiation and regulation of cell death. This review presents a brief overview of the Wnt/β-catenin signaling pathway, the regulatory mechanism of this pathway and its role in intestinal homeostasis. Additionally, this review highlights the molecular mechanisms and the histomorphological features of Wnt hyperactivation. Furthermore, the central role of the Wnt signaling pathway in intestinal carcinogenesis as well as its clinical relevance in colorectal carcinoma are discussed.

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

Abstract: Wnt signaling plays key roles in embryonic development and adult stem cell homeostasis and is altered in human cancer. 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.

Cited by 18 publications

References 80 publications

Reconstitution of the destruction complex defines roles of AXIN polymers and APC in β-catenin capture, phosphorylation, and ubiquitylation

Reconstitution of the destruction complex defines roles of AXIN polymers and APC in β-catenin capture, phosphorylation, and ubiquitylation

Multivalent tumor suppressor adenomatous polyposis coli promotes Axin biomolecular condensate formation and efficient β-catenin degradation

Intestinal Wnt in the transition from physiology to oncology

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