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Analysis of Wnt signaling β-catenin spatial dynamics in HEK293T cells
Journal article   Open access   Peer reviewed

Analysis of Wnt signaling β-catenin spatial dynamics in HEK293T cells

C. Tan, B.S. Gardiner, Y. Hirokawa, D.W. Smith and A.W. Burgess
BMC Systems Biology, Vol.8(1)
2014
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Abstract

Background Wnt/β-catenin signaling is involved in different stages of mammalian development and implicated in various cancers (e.g. colorectal cancer). Recent experimental and computational studies have revealed characteristics of the pathway, however a cell-specific spatial perspective is lacking. In this study, a novel 3D confocal quantitation protocol is developed to acquire spatial (two cellular compartments: nucleus and cytosol-membrane) and temporal quantitative data on target protein (e.g. β-catenin) concentrations in Human Epithelial Kidney cells (HEK293T) during perturbation (with either cycloheximide or Wnt3A). Computational models of the Wnt pathway are constructed and interrogated based on this data. Results A single compartment Wnt pathway model is compared with a simple β-catenin two compartment model to investigate Wnt3A signaling in HEK293T cells. When protein synthesis is inhibited, β-catenin decreases at the same rate in both cellular compartments, suggesting diffusional transport is fast compared to β-catenin degradation in the cytosol. With Wnt3A stimulation, the total amount of β-catenin rises throughout the cell, however the increase is initially (~first hour) faster in the nuclear compartment. While both models were able to reproduce the whole cell changes in β-catenin, only the compartment model reproduced the Wnt3A induced changes in β-catenin distribution and it was also the best fit for the data obtained when active transport was included alongside passive diffusion transport. Conclusions This integrated 3D quantitation imaging protocol and computational modeling approach allowed cell-specific compartment models of the signaling pathways to be constructed and analyzed. The Wnt models constructed in this study are the first for HEK293T and have suggested potential roles of inter-compartment transport to the dynamics of signaling.

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Collaboration types
Domestic collaboration
Citation topics
1 Clinical & Life Sciences
1.108 Molecular & Cell Biology - Cancer & Development
1.108.757 Wnt-Cadherin Interactions
Web Of Science research areas
Mathematical & Computational Biology
ESI research areas
Biology & Biochemistry
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