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Issue 3, 2020
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Optogenetic Rac1 engineered from membrane lipid-binding RGS-LOV for inducible lamellipodia formation

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Abstract

We report the construction of a single-component optogenetic Rac1 (opto-Rac1) to control actin polymerization by dynamic membrane recruitment. Opto-Rac1 is a fusion of wildtype human Rac1 small GTPase to the C-terminal region of BcLOV4, a LOV (light-oxygen-voltage) photoreceptor that rapidly binds the plasma membrane upon blue-light activation via a direct electrostatic interaction with anionic membrane phospholipids. Translocation of the fused wildtype Rac1 effector permits its activation by GEFs (guanine nucleotide exchange factors) and consequent actin polymerization and lamellipodia formation, unlike in existing single-chain systems that operate by allosteric photo-switching of constitutively active Rac1 or the heterodimerization-based (i.e. two-component) membrane recruitment of a Rac1-activating GEF. Opto-Rac1 induction of lamellipodia formation was spatially restricted to the patterned illumination field and was efficient, requiring sparse stimulation duty ratios of ∼1–2% (at the sensitivity threshold for flavin photocycling) to cause significant changes in cell morphology. This work exemplifies how the discovery of LOV proteins of distinct signal transmission modes can beget new classes of optogenetic tools for controlling cellular function.

Graphical abstract: Optogenetic Rac1 engineered from membrane lipid-binding RGS-LOV for inducible lamellipodia formation

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Supplementary files

Article information


Submitted
04 Nov 2019
Accepted
03 Feb 2020
First published
05 Feb 2020

Photochem. Photobiol. Sci., 2020,19, 353-361
Article type
Paper

Optogenetic Rac1 engineered from membrane lipid-binding RGS-LOV for inducible lamellipodia formation

E. E. Berlew, I. A. Kuznetsov, K. Yamada, L. J. Bugaj and B. Y. Chow, Photochem. Photobiol. Sci., 2020, 19, 353
DOI: 10.1039/C9PP00434C

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