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Spatio-Temporal Manipulation of Small GTPase Activity at Subcellular Level and on Timescale of Seconds in Living Cells
Published on: March 9, 2012
Spatiotemporal control of cell signalling using a light-switchable protein interaction
Anselm Levskaya1, Orion D Weiner, Wendell A Lim
1The Cell Propulsion Lab, UCSF/UCB NIH Nanomedicine Development Center, University of California, San Francisco, California 94158-2517, USA.
Nature
|September 15, 2009
Summary
Scientists engineered a new light-control system using plant proteins to precisely manipulate cellular behavior. This genetically encoded system allows reversible protein translocation, offering new tools for cell biology research.
Area of Science:
- Cell Biology
- Biotechnology
- Molecular Biology
Background:
- Genetically encoded optical reporters like GFP enable cellular observation.
- Optical control of cellular behavior is challenging due to difficulties in engineering light-sensitive proteins.
- Existing methods for optical control include semi-synthetic receptors and channel rhodopsins, primarily for neuronal networks.
Purpose of the Study:
- To develop a novel, genetically encoded system for precise optical control of cellular functions.
- To engineer a light-sensitive protein-protein interaction for reversible cellular manipulation.
- To demonstrate the system's utility in controlling protein localization and cell morphology.
Main Methods:
- Adapted and optimized a reversible protein-protein interaction from the Arabidopsis thaliana phytochrome signaling network.
- Engineered a genetically encoded light-control system based on this interaction.
- Utilized the system to achieve light-gated translocation of proteins to the cell membrane.
- Applied the system to control Rho-family GTPase activators to influence the actin cytoskeleton and cell morphology.
Main Results:
- Demonstrated precise and reversible translocation of target proteins to the membrane with spatial and temporal resolution (micrometer, second).
- Showcased light-gated translocation of Rho-family GTPase activators.
- Successfully reshaped and directed mammalian cell morphology using light-induced cytoskeletal changes.
- Validated the system's generic applicability for controlling diverse cellular functions.
Conclusions:
- The developed light-gated protein-protein interaction system provides a powerful new tool for optical control in cell biology.
- This system enables precise, reversible manipulation of cellular processes with high spatial and temporal resolution.
- It holds potential for creating light-programmable reagents and advancing perturbative, quantitative cell biology experiments.
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