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Concerted localization-resets precede YAP-dependent transcription

Rajarshi P. Ghosh, J. Matthew Franklin, Quanming Shi, Michael P. Reddick, Jan T. Liphardt
doi: https://doi.org/10.1101/539049
Rajarshi P. Ghosh
1Bioengineering, Stanford University, Stanford, CA 94305, USA
2BioX Institute, Stanford University, Stanford, CA 94305, USA
3ChEM-H, Stanford University, Stanford, CA 94305, USA
4Cell Biology Division, Stanford Cancer Institute, Stanford, CA 94305, USA
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  • For correspondence: rajarshi@stanford.edu jan.liphardt@stanford.edu
J. Matthew Franklin
1Bioengineering, Stanford University, Stanford, CA 94305, USA
2BioX Institute, Stanford University, Stanford, CA 94305, USA
3ChEM-H, Stanford University, Stanford, CA 94305, USA
4Cell Biology Division, Stanford Cancer Institute, Stanford, CA 94305, USA
5Chemical Engineering, Stanford University, Stanford, CA 94305, USA
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Quanming Shi
1Bioengineering, Stanford University, Stanford, CA 94305, USA
2BioX Institute, Stanford University, Stanford, CA 94305, USA
3ChEM-H, Stanford University, Stanford, CA 94305, USA
4Cell Biology Division, Stanford Cancer Institute, Stanford, CA 94305, USA
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Michael P. Reddick
1Bioengineering, Stanford University, Stanford, CA 94305, USA
2BioX Institute, Stanford University, Stanford, CA 94305, USA
3ChEM-H, Stanford University, Stanford, CA 94305, USA
4Cell Biology Division, Stanford Cancer Institute, Stanford, CA 94305, USA
5Chemical Engineering, Stanford University, Stanford, CA 94305, USA
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Jan T. Liphardt
1Bioengineering, Stanford University, Stanford, CA 94305, USA
2BioX Institute, Stanford University, Stanford, CA 94305, USA
3ChEM-H, Stanford University, Stanford, CA 94305, USA
4Cell Biology Division, Stanford Cancer Institute, Stanford, CA 94305, USA
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  • For correspondence: rajarshi@stanford.edu jan.liphardt@stanford.edu
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Abstract

Yes-associated protein 1 (YAP) is a transcriptional regulator with critical roles in mechanotransduction, organ size control, and regeneration. Here, using new tools for real-time visualization of native YAP and target gene transcription dynamics, we show that a cycle of fast exodus of nuclear YAP to the cytoplasm followed by fast reentry to the nucleus (“localization-resets”) activates YAP target genes. These “resets” could be induced by calcium signaling, modulation of actomyosin contractility, or mitosis. Using nascent-transcription reporter knock-ins of YAP target genes, we observed a strict association between these resets and downstream transcription. Oncogenically-transformed transformed cell lines lacked localization-resets and instead showed dramatically elevated rates of nucleocytoplasmic shuttling of YAP, suggesting an escape from compartmentalization-based control. The single-cell localization and transcription traces suggest that YAP activity is not a simple linear function of nuclear enrichment and point to a new model of transcriptional activation based on nucleocytoplasmic exchange properties of YAP.

Footnotes

  • The manuscript has been modified considerably with a host of new experiments and also a strategic redistribution of earlier figure components to help ease the flow of the paper.

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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. All rights reserved. No reuse allowed without permission.
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Posted April 05, 2020.
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Concerted localization-resets precede YAP-dependent transcription
Rajarshi P. Ghosh, J. Matthew Franklin, Quanming Shi, Michael P. Reddick, Jan T. Liphardt
bioRxiv 539049; doi: https://doi.org/10.1101/539049
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Concerted localization-resets precede YAP-dependent transcription
Rajarshi P. Ghosh, J. Matthew Franklin, Quanming Shi, Michael P. Reddick, Jan T. Liphardt
bioRxiv 539049; doi: https://doi.org/10.1101/539049

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