Extraction of active RhoGTPases by RhoGDI regulates spatiotemporal patterning of RhoGTPases
- University of Wisconsin-Madison, United States
- Max Planck Institute of Molecular Physiology, Germany
Abstract
The RhoGTPases are characterized as membrane-associated molecular switches that cycle between active, GTP-bound and inactive, GDP-bound states. However, 90-95% of RhoGTPases are maintained in a soluble form by RhoGDI, which is generally viewed as a passive shuttle for inactive RhoGTPases. Our current understanding of RhoGTPase:RhoGDI dynamics has been limited by two experimental challenges: direct visualization of the RhoGTPases in vivo and reconstitution of the cycle in vitro. We developed methods to directly image vertebrate RhoGTPases in vivo or on lipid bilayers in vitro. Using these methods, we identified pools of active and inactive RhoGTPase associated with the membrane, found that RhoGDI can extract both inactive and active RhoGTPases, and found that extraction of active RhoGTPase contributes to their spatial regulation around cell wounds. These results indicate that RhoGDI directly contributes to the spatiotemporal patterning of RhoGTPases by removing active RhoGTPases from the plasma membrane.
Data availability
All quantifications made in this study are included as source data files by figure.
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Author details
Funding
National Institutes of Health (GM52932)
- William M Bement
University of Wisconsin-Madison (Dr. Stanley and Dr. Eva Lurie Weinreb Fellowship)
- Adriana E Golding
Human Frontier Science Program (HSFP CDA00070-2017-2)
- Peter Bieling
Max Planck Society (MaxSynBio Consortium)
- Ilaria Visco
The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.
Ethics
Animal experimentation: The University of Wisconsin-Madison Animal Care and Use Committee has reviewed and approved all of the experiments performed for this study, outlined in protocol G005386-RO1.
Copyright
© 2019, Golding et al.
This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.
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Extraction of active RhoGTPases by RhoGDI regulates spatiotemporal patterning of RhoGTPases
eLife 8:e50471.
https://doi.org/10.7554/eLife.50471
