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Quantitative live-cell imaging yields novel insight into endogenous WNT/CTNNB1 signaling dynamics

View ORCID ProfileS.M.A. de Man, G. Zwanenburg, M.A. Hink, View ORCID ProfileR. van Amerongen
doi: https://doi.org/10.1101/2020.05.28.120543
S.M.A. de Man
1Swammerdam Institute for Life Sciences, Section of Molecular Cytology, University of Amsterdam, Amsterdam, the Netherlands
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  • ORCID record for S.M.A. de Man
G. Zwanenburg
2Swammerdam Institute for Life Sciences, Biosystems Data Analysis Group, University of Amsterdam, Amsterdam, The Netherlands
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  • For correspondence: r.vanamerongen@uva.nl m.a.hink@uva.nl g.zwanenburg@uva.nl
M.A. Hink
1Swammerdam Institute for Life Sciences, Section of Molecular Cytology, University of Amsterdam, Amsterdam, the Netherlands
3Swammerdam Institute for Life Sciences, van Leeuwenhoek Centre for Advanced Microscopy, University of Amsterdam, Amsterdam, the Netherlands
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  • For correspondence: r.vanamerongen@uva.nl m.a.hink@uva.nl g.zwanenburg@uva.nl
R. van Amerongen
1Swammerdam Institute for Life Sciences, Section of Molecular Cytology, University of Amsterdam, Amsterdam, the Netherlands
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  • For correspondence: r.vanamerongen@uva.nl m.a.hink@uva.nl g.zwanenburg@uva.nl
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Abstract

WNT/CTNNB1 signaling regulates tissue development and homeostasis in all multicellular animals. Multiple aspects of the underlying molecular mechanism remain poorly understood and critical information on endogenous WNT/CTNNB1 signaling dynamics is missing. Here we combine CRISPR/Cas9-mediated genome editing and quantitative live-cell microscopy to measure diffusion characteristics of fluorescently tagged, endogenous CTNNB1 in human cells with high spatiotemporal resolution. State-of-the-art functional imaging reveals that both in the absence and presence of WNT, a substantial fraction of CTNNB1 resides in slow-diffusing complexes in the cytoplasm and that WNT stimulation changes their identity. We also measure the concentration of complexed and free CTNNB1 in both the cytoplasm and the nucleus before and after WNT stimulation, and use these parameters to build a minimal computational model of WNT/CTNNB1 signaling. Our work reveals that WNT regulates the dynamic distribution of CTNNB1 across different functional pools by modulating the destruction complex, nucleocytoplasmic shuttling and nuclear retention.

Competing Interest Statement

The authors have declared no competing interest.

Footnotes

  • ↵# Twitter: @wntlab

  • https://osf.io/dczx8/

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The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY-NC 4.0 International license.
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Posted May 28, 2020.
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Quantitative live-cell imaging yields novel insight into endogenous WNT/CTNNB1 signaling dynamics
S.M.A. de Man, G. Zwanenburg, M.A. Hink, R. van Amerongen
bioRxiv 2020.05.28.120543; doi: https://doi.org/10.1101/2020.05.28.120543
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Quantitative live-cell imaging yields novel insight into endogenous WNT/CTNNB1 signaling dynamics
S.M.A. de Man, G. Zwanenburg, M.A. Hink, R. van Amerongen
bioRxiv 2020.05.28.120543; doi: https://doi.org/10.1101/2020.05.28.120543

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