Crystal structure of a mammalian Wnt–frizzled complex

Hirai, Haruhiko; Matoba, Kyoko; Mihara, Emiko; Arimori, Takao; Takagi, Junichi

doi:10.1038/s41594-019-0216-z

Cited by 103 publications

(142 citation statements)

References 36 publications

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“…The palmitoleic moiety is also required for direct binding sites for Wnt ligand and its receptor Frizzled on the cell membrane (Fig. 1, C and D) (Janda et al, 2012;Hirai et al, 2019). Previous studies from our group have shown that all assessable human Wnts lost signaling activity when PORCN was knocked out, and this could be rescued by PORCN re-expression (Najdi et al, 2012).…”

Section: Wnt Signaling Pathwaymentioning

confidence: 89%

“…The crystal structure of Xenopus WNT8 in complex with mouse Frizzled-8 CRD reveals the multiple interacting surfaces of Wnt-Fzd binding, including a hydrophobic groove in the CRD that binds to the hydrophobic palmitoleate on Wnt (Janda et al, 2012). Additional structures have extended these results, suggesting palmitoleate binding serves to dimerize Frizzled CRDs (Hirai et al, 2019;Nile and Hannoush, 2019).…”

Section: Wnt Signaling Pathwaymentioning

confidence: 90%

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Wnt Signaling and Drug Resistance in Cancer

2019

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Wnts are secreted proteins that bind to cell surface receptors to activate downstream signaling cascades. Normal Wnt signaling plays key roles in embryonic development and adult tissue homeostasis. The secretion of Wnt ligands, the turnover of Wnt receptors, and the signaling transduction are tightly regulated and fine-tuned to keep the signaling output "just right." Hyperactivated Wnt signaling due to recurrent genetic alterations drives several human cancers. Elevated Wnt signaling also confers resistance to multiple conventional and targeted cancer therapies through diverse mechanisms including maintaining the cancer stem cell population, enhancing DNA damage repair, facilitating transcriptional plasticity, and promoting immune evasion. Different classes of Wnt signaling inhibitors targeting key nodes of the pathway have been developed and show efficacy in treating Wnt-driven cancers and subverting Wnt-mediated therapy resistance in preclinical studies. Several of these inhibitors have advanced to clinical trials, both singly and in combination with other existing US Food and Drug Administration-approved anti-cancer modalities. In the near future, pharmacological inhibition of Wnt signaling may be a real choice for patients with cancer. SIGNIFICANCE STATEMENTThe latest insights in Wnt signaling, ranging from basic biology to therapeutic implications in cancer, are reviewed. Recent studies extend understanding of this ancient signaling pathway and describe the development and improvement of anti-Wnt therapeutic modalities for cancer.

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Section: Wnt Signaling Pathwaymentioning

confidence: 89%

Section: Wnt Signaling Pathwaymentioning

confidence: 90%

Wnt Signaling and Drug Resistance in Cancer

2019

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“…Encouraged by recent reports of successful tagging of mouse Wnt-3a (16-18), we generated eGFP-Wnt-3a with the aim of utilizing this for ligand-receptor interaction studies. We fused eGFP directly to the N terminus of Wnt-3a, which projects away from the Wnt/FZD-CRD binding region (21,22), using a short peptide linker ( Fig. 1a and Fig.…”

Section: Characterization Of a Functionally Active Egfp-wnt3amentioning

confidence: 99%

eGFP-tagged Wnt-3a enables functional analysis of Wnt trafficking and signaling and kinetic assessment of Wnt binding to full-length Frizzled

Wesslowski

Kozielewicz

Wang

et al. 2020

Journal of Biological Chemistry

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The Wingless/Int1 (Wnt) signaling system plays multiple, essential roles in embryonic development, tissue homeostasis, and human diseases. Although many of the underlying signaling mechanisms are becoming clearer, the binding mode, kinetics, and selectivity of 19 mammalian WNTs to their receptors of the class Frizzled (FZD1–10) remain obscure. Attempts to investigate Wnt-FZD interactions are hampered by the difficulties in working with Wnt proteins and their recalcitrance to epitope tagging. Here, we used a fluorescently tagged version of mouse Wnt-3a for studying Wnt-FZD interactions. We observed that the enhanced GFP (eGFP)-tagged Wnt-3a maintains properties akin to wild-type (WT) Wnt-3a in several biologically relevant contexts. The eGFP-tagged Wnt-3a was secreted in an evenness interrupted (EVI)/Wntless-dependent manner, activated Wnt/β-catenin signaling in 2D and 3D cell culture experiments, promoted axis duplication in Xenopus embryos, stimulated low-density lipoprotein receptor-related protein 6 (LRP6) phosphorylation in cells, and associated with exosomes. Further, we used conditioned medium containing eGFP-Wnt-3a to visualize its binding to FZD and to quantify Wnt-FZD interactions in real time in live cells, utilizing a recently established NanoBRET-based ligand binding assay. In summary, the development of a biologically active, fluorescent Wnt-3a reported here opens up the technical possibilities to unravel the intricate biology of Wnt signaling and Wnt-receptor selectivity.

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“…However, while in Zebrafish models its importance is more evident (Jing et al, 2009) the corresponding mouse models have provided eloquent but less obvious results (Messeant et al, 2015;Remedio et al, 2016). Nonetheless, a structural comparison of the rat MuSK Fz-CRD (Stiegler et al, 2009) with the mouse Wnt3-Frizzled8 complex (Hirai et al, 2019) seems to define a clearer picture. This shows how the MuSK Fz-CRD might be conductive to Wnt binding by accommodating the diametrically opposed binding of the Wnt palmitate tail and Zn-finger domain (Figure 7A) much like the already available structures of Wnt:Frizzled complexes (Janda et al, 2012;Hirai et al, 2019).…”

Section: The "Structural" Possibility Of Wnt-lrp4:musk Cross-talkmentioning

confidence: 99%

“…Nonetheless, a structural comparison of the rat MuSK Fz-CRD (Stiegler et al, 2009) with the mouse Wnt3-Frizzled8 complex (Hirai et al, 2019) seems to define a clearer picture. This shows how the MuSK Fz-CRD might be conductive to Wnt binding by accommodating the diametrically opposed binding of the Wnt palmitate tail and Zn-finger domain (Figure 7A) much like the already available structures of Wnt:Frizzled complexes (Janda et al, 2012;Hirai et al, 2019). Interestingly, the MuSK Fz-CRD adopts two distinct conformations in crystal structures (Stiegler et al, 2009), respectively likely or unlikely to bind Wnts (Figure 7B).…”

Section: The "Structural" Possibility Of Wnt-lrp4:musk Cross-talkmentioning

confidence: 99%

Dissecting the Extracellular Complexity of Neuromuscular Junction Organizers

Guarino

Canciani

Forneris

2020

Front. Mol. Biosci.

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Synapse formation is a very elaborate process dependent upon accurate coordination of pre and post-synaptic specialization, requiring multiple steps and a variety of receptors and signaling molecules. Due to its relative structural simplicity and the ease in manipulation and observation, the neuromuscular synapse or neuromuscular junction (NMJ)-the connection between motor neurons and skeletal muscle-represents the archetype junction system for studying synapse formation and conservation. This junction is essential for survival, as it controls our ability to move and breath. NMJ formation requires coordinated interactions between motor neurons and muscle fibers, which ultimately result in the formation of a highly specialized post-synaptic architecture and a highly differentiated nerve terminal. Furthermore, to ensure a fast and reliable synaptic transmission following neurotransmitter release, ligand-gated channels (acetylcholine receptors, AChRs) are clustered on the post-synaptic muscle cell at high concentrations in sites opposite the presynaptic active zone, supporting a direct role for nerves in the organization of the post-synaptic membrane architecture. This organized clustering process, essential for NMJ formation and for life, relies on key signaling molecules and receptors and is regulated by soluble extracellular molecules localized within the synaptic cleft. Notably, several mutations as well as auto-antibodies against components of these signaling complexes have been related to neuromuscular disorders. The recent years have witnessed strong progress in the understanding of molecular identities, architectures, and functions of NMJ macromolecules. Among these, prominent roles have been proposed for neural variants of the proteoglycan agrin, its receptor at NMJs composed of the lipoprotein receptor-related protein 4 (LRP4) and the muscle-specific kinase (MuSK), as well as the regulatory soluble synapse-specific protease Neurotrypsin. In this review we summarize the current state of the art regarding molecular structures and (agrin-dependent) canonical, as well as (agrin-independent) non-canonical, MuSK signaling mechanisms that underscore the formation of neuromuscular junctions, with the aim of providing a broad perspective to further stimulate molecular, cellular and tissue biology investigations on this fundamental intercellular contact.

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Crystal structure of a mammalian Wnt–frizzled complex

Cited by 103 publications

References 36 publications

Wnt Signaling and Drug Resistance in Cancer

Wnt Signaling and Drug Resistance in Cancer

eGFP-tagged Wnt-3a enables functional analysis of Wnt trafficking and signaling and kinetic assessment of Wnt binding to full-length Frizzled

Dissecting the Extracellular Complexity of Neuromuscular Junction Organizers

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