Quantitative fluorescence microscopy for the analysis of Wnt pathway interactions and dynamics
Funding period: 2017-2025
Project Leaders
Abstract
In this collaborative project the quantification of Wnt ligand interactions will be extended to all ten FZD receptors, thereby gaining important, unresolved information on the native binding selectivity of a Wnt and its potential to dictate Wnt signaling specificity. FFS approaches (e.g., axial line-scanning FCS) will be used to quantify ligand-receptor and receptor-receptor interactions involved in signalosome formation. By super-resolution single-molecule localization microscopy imaging (SMLM) the stoichiometry and dynamics of Wnt receptor/co-receptor clusters and signalosome formation at the plasma membrane will be visualized. The project relies on both novel technologies and interdisciplinary collaborations within the consortium.
Project-related Publications
NanoBiT- and NanoBiT/BRET-based assays allow the analysis of binding kinetics of WNT-3A to endogenous Frizzled 7 in a colorectal cancer model
N-Glycosylation of LRP6 by B3GnT2 Promotes Wnt/β-Catenin Signalling
Allele-specific endogenous tagging and quantitative analysis of β-catenin in colorectal cancer cells
Two plus one is almost three: a fast approximation for multi-view deconvolution
Biomed Opt Express. 2021 Dec 7;13(1):147-158. doi: 10.1364/BOE.443660. eCollection 2022 Jan 1. PMID: 35154860
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Davidson G. Handb Exp Pharmacol. 2021;269:45-73. doi: 10.1007/164_2021_526. PMID: 34490514
Learn moreQuantitative Profiling of WNT-3A Binding to All Human Frizzled Paralogues in HEK293 Cells by NanoBiT/BRET Assessments
ACS Pharmacol Transl Sci. 2021 May 11;4(3):1235-1245. doi: 10.1021/acsptsci.1c00084. eCollection 2021 Jun 11. PMID: 34151213
Learn moreSuper-resolution RNA imaging using a rhodamine-binding aptamer with fast exchange kinetics
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eGFP-tagged Wnt-3a enables functional analysis of Wnt trafficking and signaling and kinetic assessment of Wnt binding to full-length Frizzled
Wesslowski J, Kozielewicz P, Wang X, Cui H, Schihada H, Kranz D, Karuna M P, Levkin P, Gross JC, Boutros M, Schulte G, Davidson G. J Biol Chem. 2020 May 7. pii: jbc.RA120.012892. PMID:32381507
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Learn moreLef1 regulates caveolin expression and caveolin dependent endocytosis, a process necessary for Wnt5a/Ror2 signaling during Xenopus gastrulation
Sci Rep. 2019 Oct 30;9(1):15645. doi: 10.1038/s41598-019-52218-1. PMID: 31666627
Learn moreWnt/PCP controls spreading of Wnt/β-catenin signals by cytonemes in vertebrates
Mattes B, Dang Y, Greicius G, Kaufmann LT, Prunsche B, Rosenbauer J, Stegmaier J, Mikut R, Özbek S, Nienhaus GU, Schug A, Virshup DM, Scholpp S. Elife. 2018 Jul 31;7. PMID:30060804
Learn moreBackground suppression in fluorescence nanoscopy with stimulated emission double depletion
Gao, P., B. Prunsche, L. Zhou, K. Nienhaus, and G.U. Nienhaus. 2017. Nat Photon. 11:163-169.
Learn moreDual-color dual-focus line-scanning FCS for quantitative analysis of receptor-ligand interactions in living specimens
Sci Rep. 2015 May 7;5:10149. doi: 10.1038/srep10149. PMID: 25951521
Learn moreTyrosine phosphorylation of LRP6 by Src and Fer inhibits Wnt/β-catenin signaling
EMBO Rep. 2014 Dec;15(12):1254-67. doi: 10.15252/embr.201439644. Epub 2014 Nov 12. PMID: 25391905
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