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Endogenous Bioid Elucidates TCF7L1 Interactome Modulation Upon GSK-3 Inhibition in Mouse ESCs

19 Pages Posted: 7 Mar 2019 Sneak Peek Status: Under Review

See all articles by Steven Moreira

Steven Moreira

McMaster University - Department of Biochemistry and Biomedical Sciences

Caleb Seo

McMaster University - Department of Biochemistry and Biomedical Sciences

Victor Gordon

McMaster University - Department of Biochemistry and Biomedical Sciences

Sansi Xing

McMaster University - Department of Biochemistry and Biomedical Sciences

Ruilin Wu

McMaster University - Department of Biochemistry and Biomedical Sciences

Enio Polena

McMaster University - Department of Biochemistry and Biomedical Sciences

Vincent Fung

McMaster University - Department of Biochemistry and Biomedical Sciences

Deborah Ng

University Health Network - Princess Margaret Cancer Centre; University of Toronto - Department of Medical Biophysics

Cassandra J. Wong

University of Toronto - Lunenfeld-Tanenbaum Research Institute; University of Toronto - Department of Molecular Genetics

Brett Larsen

University of Toronto - Lunenfeld-Tanenbaum Research Institute; University of Toronto - Department of Molecular Genetics

Brian Raught

University Health Network - Princess Margaret Cancer Centre; University of Toronto - Department of Medical Biophysics

Anne-Claude Gingras

University of Toronto - Lunenfeld-Tanenbaum Research Institute; University of Toronto - Department of Molecular Genetics

Yu Lu

McMaster University - Department of Biochemistry and Biomedical Sciences

Bradley W. Doble

McMaster University - Department of Biochemistry and Biomedical Sciences

More...

Abstract

Modulation of Wnt target gene expression via the TCF/LEFs remains poorly understood. We employ proximity-based biotin labeling (BioID) to examine GSK-3 inhibitor effects on the TCF7L1 interactome in mouse ESCs. We generated ESC lines with biotin ligase BirA* fused to TCF7L1 by knocking it into the endogenous TCF7L1 locus or by inserting a dox-inducible BirA*-TCF7L1 transgene into the Rosa26 locus. Induction yielded BirA*-TCF7L1 levels 3-fold higher than in the endogenous system, but substantial overlap in biotinylated proteins with high peptide counts were detected by each method. Known TCF7L1 interactors TLE3/4 and β-catenin, and numerous proteins not previously associated with TCF7L1, were identified in both systems. Despite reduced BirA*-TCF7L1 levels, the number of hits identified with both BioID approaches increased after GSK-3 inhibition. We elucidate the network of TCF7L1 proximal proteins regulated by GSK-3 inhibition, validate the utility of endogenous BioID, and provide mechanistic insights into TCF7L1 target gene regulation.

Suggested Citation

Moreira, Steven and Seo, Caleb and Gordon, Victor and Xing, Sansi and Wu, Ruilin and Polena, Enio and Fung, Vincent and Ng, Deborah and Wong, Cassandra J. and Larsen, Brett and Raught, Brian and Gingras, Anne-Claude and Lu, Yu and Doble, Bradley W., Endogenous Bioid Elucidates TCF7L1 Interactome Modulation Upon GSK-3 Inhibition in Mouse ESCs (March 7, 2019). Available at SSRN: https://ssrn.com/abstract=3348349 or http://dx.doi.org/10.2139/ssrn.3348349
This is a paper under consideration at Cell Press and has not been peer-reviewed.

Steven Moreira

McMaster University - Department of Biochemistry and Biomedical Sciences

1280 Main Street West
Hamilton, Ontario L8S 4L8
Canada

Caleb Seo

McMaster University - Department of Biochemistry and Biomedical Sciences

1280 Main Street West
Hamilton, Ontario L8S 4L8
Canada

Victor Gordon

McMaster University - Department of Biochemistry and Biomedical Sciences

1280 Main Street West
Hamilton, Ontario L8S 4L8
Canada

Sansi Xing

McMaster University - Department of Biochemistry and Biomedical Sciences

1280 Main Street West
Hamilton, Ontario L8S 4L8
Canada

Ruilin Wu

McMaster University - Department of Biochemistry and Biomedical Sciences

1280 Main Street West
Hamilton, Ontario L8S 4L8
Canada

Enio Polena

McMaster University - Department of Biochemistry and Biomedical Sciences

1280 Main Street West
Hamilton, Ontario L8S 4L8
Canada

Vincent Fung

McMaster University - Department of Biochemistry and Biomedical Sciences

1280 Main Street West
Hamilton, Ontario L8S 4L8
Canada

Deborah Ng

University Health Network - Princess Margaret Cancer Centre

610 University Avenue
Toronto, ON M5G 2M9
Canada

University of Toronto - Department of Medical Biophysics

Toronto, Ontario M5S 3G8
Canada

Cassandra J. Wong

University of Toronto - Lunenfeld-Tanenbaum Research Institute

600 University Ave
Toronto, Ontario
Canada

University of Toronto - Department of Molecular Genetics

Toronto, Ontario
Canada

Brett Larsen

University of Toronto - Lunenfeld-Tanenbaum Research Institute

600 University Ave
Toronto, Ontario
Canada

University of Toronto - Department of Molecular Genetics

Toronto, Ontario
Canada

Brian Raught

University Health Network - Princess Margaret Cancer Centre

610 University Avenue
Toronto, ON M5G 2M9
Canada

University of Toronto - Department of Medical Biophysics

Toronto, Ontario M5S 3G8
Canada

Anne-Claude Gingras

University of Toronto - Lunenfeld-Tanenbaum Research Institute

600 University Ave
Toronto, Ontario
Canada

University of Toronto - Department of Molecular Genetics

Toronto, Ontario
Canada

Yu Lu

McMaster University - Department of Biochemistry and Biomedical Sciences

1280 Main Street West
Hamilton, Ontario L8S 4L8
Canada

Bradley W. Doble (Contact Author)

McMaster University - Department of Biochemistry and Biomedical Sciences ( email )

1280 Main Street West
Hamilton, Ontario L8S 4L8
Canada

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