Wnt-inducible Lrp6-APEX2 interacting proteins identify ESCRT machinery and Trk-fused gene as components of the Wnt signaling pathway

Abstract The canonical Wnt pathway serves as a hub connecting diverse cellular processes, including β-catenin signaling, differentiation, growth, protein stability, macropinocytosis, and nutrient acquisition in lysosomes. We have proposed that sequestration of β-catenin destruction complex component...

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Autores principales:	Gabriele Colozza, Yasaman Jami-Alahmadi, Alyssa Dsouza, Nydia Tejeda-Muñoz, Lauren V. Albrecht, Eric A. Sosa, James A. Wohlschlegel, Edward M. De Robertis
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2020
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/70bc8e647cea4e6a97568c3d15403f70
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spelling	oai:doaj.org-article:70bc8e647cea4e6a97568c3d15403f702021-12-02T12:33:14ZWnt-inducible Lrp6-APEX2 interacting proteins identify ESCRT machinery and Trk-fused gene as components of the Wnt signaling pathway10.1038/s41598-020-78019-52045-2322https://doaj.org/article/70bc8e647cea4e6a97568c3d15403f702020-12-01T00:00:00Zhttps://doi.org/10.1038/s41598-020-78019-5https://doaj.org/toc/2045-2322Abstract The canonical Wnt pathway serves as a hub connecting diverse cellular processes, including β-catenin signaling, differentiation, growth, protein stability, macropinocytosis, and nutrient acquisition in lysosomes. We have proposed that sequestration of β-catenin destruction complex components in multivesicular bodies (MVBs) is required for sustained canonical Wnt signaling. In this study, we investigated the events that follow activation of the canonical Wnt receptor Lrp6 using an APEX2-mediated proximity labeling approach. The Wnt co-receptor Lrp6 was fused to APEX2 and used to biotinylate targets that are recruited near the receptor during Wnt signaling at different time periods. Lrp6 proximity targets were identified by mass spectrometry, and revealed that many endosomal proteins interacted with Lrp6 within 5 min of Wnt3a treatment. Interestingly, we found that Trk-fused gene (TFG), previously known to regulate the cell secretory pathway and to be rearranged in thyroid and lung cancers, was strongly enriched in the proximity of Lrp6. TFG depletion with siRNA, or knock-out with CRISPR/Cas9, significantly reduced Wnt/β-catenin signaling in cell culture. In vivo, studies in the Xenopus system showed that TFG is required for endogenous Wnt-dependent embryonic patterning. The results suggest that the multivesicular endosomal machinery and the novel player TFG have important roles in Wnt signaling.Gabriele ColozzaYasaman Jami-AlahmadiAlyssa DsouzaNydia Tejeda-MuñozLauren V. AlbrechtEric A. SosaJames A. WohlschlegelEdward M. De RobertisNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 10, Iss 1, Pp 1-14 (2020)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Gabriele Colozza Yasaman Jami-Alahmadi Alyssa Dsouza Nydia Tejeda-Muñoz Lauren V. Albrecht Eric A. Sosa James A. Wohlschlegel Edward M. De Robertis Wnt-inducible Lrp6-APEX2 interacting proteins identify ESCRT machinery and Trk-fused gene as components of the Wnt signaling pathway
description	Abstract The canonical Wnt pathway serves as a hub connecting diverse cellular processes, including β-catenin signaling, differentiation, growth, protein stability, macropinocytosis, and nutrient acquisition in lysosomes. We have proposed that sequestration of β-catenin destruction complex components in multivesicular bodies (MVBs) is required for sustained canonical Wnt signaling. In this study, we investigated the events that follow activation of the canonical Wnt receptor Lrp6 using an APEX2-mediated proximity labeling approach. The Wnt co-receptor Lrp6 was fused to APEX2 and used to biotinylate targets that are recruited near the receptor during Wnt signaling at different time periods. Lrp6 proximity targets were identified by mass spectrometry, and revealed that many endosomal proteins interacted with Lrp6 within 5 min of Wnt3a treatment. Interestingly, we found that Trk-fused gene (TFG), previously known to regulate the cell secretory pathway and to be rearranged in thyroid and lung cancers, was strongly enriched in the proximity of Lrp6. TFG depletion with siRNA, or knock-out with CRISPR/Cas9, significantly reduced Wnt/β-catenin signaling in cell culture. In vivo, studies in the Xenopus system showed that TFG is required for endogenous Wnt-dependent embryonic patterning. The results suggest that the multivesicular endosomal machinery and the novel player TFG have important roles in Wnt signaling.
format	article
author	Gabriele Colozza Yasaman Jami-Alahmadi Alyssa Dsouza Nydia Tejeda-Muñoz Lauren V. Albrecht Eric A. Sosa James A. Wohlschlegel Edward M. De Robertis
author_facet	Gabriele Colozza Yasaman Jami-Alahmadi Alyssa Dsouza Nydia Tejeda-Muñoz Lauren V. Albrecht Eric A. Sosa James A. Wohlschlegel Edward M. De Robertis
author_sort	Gabriele Colozza
title	Wnt-inducible Lrp6-APEX2 interacting proteins identify ESCRT machinery and Trk-fused gene as components of the Wnt signaling pathway
title_short	Wnt-inducible Lrp6-APEX2 interacting proteins identify ESCRT machinery and Trk-fused gene as components of the Wnt signaling pathway
title_full	Wnt-inducible Lrp6-APEX2 interacting proteins identify ESCRT machinery and Trk-fused gene as components of the Wnt signaling pathway
title_fullStr	Wnt-inducible Lrp6-APEX2 interacting proteins identify ESCRT machinery and Trk-fused gene as components of the Wnt signaling pathway
title_full_unstemmed	Wnt-inducible Lrp6-APEX2 interacting proteins identify ESCRT machinery and Trk-fused gene as components of the Wnt signaling pathway
title_sort	wnt-inducible lrp6-apex2 interacting proteins identify escrt machinery and trk-fused gene as components of the wnt signaling pathway
publisher	Nature Portfolio
publishDate	2020
url	https://doaj.org/article/70bc8e647cea4e6a97568c3d15403f70
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Wnt-inducible Lrp6-APEX2 interacting proteins identify ESCRT machinery and Trk-fused gene as components of the Wnt signaling pathway

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