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Live Imaging of Xwnt5A-ROR2 Complexes

Wallkamm, V. 1; Dörlich, R. 2,3; Rahm, K. 1; Klessing, T. 1; Nienhaus, G. U. ORCID iD icon 2,3; Wedlich, D. 1; Gradl, D. 1
1 Zoologisches Institut (ZOO), Karlsruher Institut für Technologie (KIT)
2 Institut für Angewandte Physik (APH), Karlsruher Institut für Technologie (KIT)
3 Institut für Toxikologie und Genetik (ITG), Karlsruher Institut für Technologie (KIT)

Abstract:

Secreted molecules of the Wnt family regulate key decisions in embryogenesis and adult tissue homeostasis by activating a complex network of Wnt signaling pathways. Although the different branches of Wnt signaling have been studied for more than 25 years, fluorophore tagged constructs for live cell imaging of Wnt molecules activating the Wnt/β-catenin pathway have become available only recently. We have generated a fluorophore tagged Wnt construct of the Xenopus Wnt5a protein (Xwnt5A) with the enhanced green fluorescent protein (EGFP), Xwnt5A-EGFP. This construct activates non-canonical Wnt pathways in an endocytosis dependent manner and is capable of compensating for the loss of endogenous Xwnt5A in Xenopus embryos. Strikingly, non-canonical Wnt pathway activation was restricted to short-range signaling while an inhibitory effect was observed in transwell cell cultures taken as long-range signaling model sytem. We used our Xwnt5A-EGFP construct to analyze in vivo binding of Wnt5A to its co-receptor ROR2 on the microscopic and on the molecular level. On the microscopic level, Xwnt5A-EGFP clusters in the membrane and recruits ROR2-mCherry to these clusters. ... mehr


Volltext §
DOI: 10.5445/IR/1000043637
Originalveröffentlichung
DOI: 10.1371/journal.pone.0109428
Scopus
Zitationen: 15
Web of Science
Zitationen: 14
Dimensions
Zitationen: 20
Cover der Publikation
Zugehörige Institution(en) am KIT Zoologisches Institut (ZOO)
Publikationstyp Zeitschriftenaufsatz
Publikationsjahr 2014
Sprache Englisch
Identifikator ISSN: 1932-6203
urn:nbn:de:swb:90-436377
KITopen-ID: 1000043637
Erschienen in PLoS One
Verlag Public Library of Science (PLoS)
Band 9
Heft 10
Seiten e109428
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