A06

Quantitative fluorescence microscopy for the analysis of Wnt pathway interactions and dynamics

Funding period: 2017-2025

Project A06

Project Leaders

Dr. Gary Davidson

Dr. Gary Davidson

Project Leader A06
Prof. Dr. Gerd Ulrich Nienhaus

Prof. Dr. Gerd Ulrich Nienhaus

Project Leader A06, Z02

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

Allele-specific endogenous tagging and quantitative analysis of β-catenin in colorectal cancer cells

Ambrosi G, Voloshanenko O, Eckert AF, Kranz D, Nienhaus GU, Boutros M. Elife. 2022 Jan 11;11:e64498. doi: 10.7554/eLife.64498.
PMID: 35014953

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Two plus one is almost three: a fast approximation for multi-view deconvolution

Hüpfel M, Fernández Merino M, Bennemann J, Takamiya M, Rastegar S, Tursch A, Holstein TW, Nienhaus GU. 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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LRPs in WNT Signalling

Davidson G. (2021) LRPs in WNT Signalling. In: Schulte G., Kozielewicz P. (eds) Pharmacology of the WNT Signaling System. Handbook of Experimental Pharmacology, vol 269. Springer, Cham.  

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Quantitative Profiling of WNT-3A Binding to All Human Frizzled Paralogues in HEK293 Cells by NanoBiT/BRET Assessments

Kozielewicz P, Shekhani R, Moser S, Bowin CF, Wesslowski J, Davidson G, Schulte G. ACS Pharmacol Transl Sci. 2021 May 11;4(3):1235-1245. doi: 10.1021/acsptsci.1c00084. eCollection 2021 Jun 11. PMID: 34151213

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Super-resolution RNA imaging using a rhodamine-binding aptamer with fast exchange kinetics

Sunbul M, Lackner J, Martin A, Englert D, Hacene B, Grün F, Nienhaus K, Nienhaus GU, Jäschke A. Nat Biotechnol. 2021 Jun;39(6):686-690. doi: 10.1038/s41587-020-00794-3. Epub 2021 Feb 11.
PMID: 33574610

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Measuring ligand-cell surface receptor affinities with axial line-scanning fluorescence correlation spectroscopy

Eckert AF, Gao P, Wesslowski J, Wang X, Rath J, Nienhaus K, Davidson G, Nienhaus GU. Elife. 2020 May 22;9:e55286.

PMID: 32441251

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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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Fam83F induces p53 stabilisation and promotes its activity

Salama M, Benitez-Riquelme D, Elabd S, Munoz L, Zhang P, Glanemann M, Mione MC, Goldin R, Soussi T, Davidson G, Blattner C. Cell Death Differ. 2019 Oct;26(10):2125-2138. doi: 10.1038/s41418-019-0281-1. Epub 2019 Jan 28. PMID: 30692643

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Lef1 regulates caveolin expression and caveolin dependent endocytosis, a process necessary for Wnt5a/Ror2 signaling during Xenopus gastrulation

Puzik K, Tonnier V, Opper I, Eckert A, Zhou L, Kratzer MC, Noble FL, Nienhaus GU, Gradl D. Sci Rep. 2019 Oct 30;9(1):15645. doi: 10.1038/s41598-019-52218-1. PMID: 31666627

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Wnt/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

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Background 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.

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Dual-color dual-focus line-scanning FCS for quantitative analysis of receptor-ligand interactions in living specimens

Dörlich RM, Chen Q, Niklas Hedde P, Schuster V, Hippler M, Wesslowski J, Davidson G, Nienhaus GU. Sci Rep. 2015 May 7;5:10149. doi: 10.1038/srep10149. PMID: 25951521

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Tyrosine phosphorylation of LRP6 by Src and Fer inhibits Wnt/β-catenin signaling

Chen Q, Su Y, Wesslowski J, Hagemann AI, Ramialison M, Wittbrodt J, Scholpp S, Davidson G. EMBO Rep. 2014 Dec;15(12):1254-67. doi: 10.15252/embr.201439644. Epub 2014 Nov 12. PMID: 25391905

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Wnt induces LRP6 signalosomes and promotes dishevelled-dependent LRP6 phosphorylation

Bilic J, Huang YL, Davidson G, Zimmermann T, Cruciat CM, Bienz M, Niehrs C. Science. 2007 Jun 15;316(5831):1619-22. doi: 10.1126/science.1137065.
PMID: 17569865

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