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Single‐Component Optogenetic Tools for Inducible RhoA GTPase Signaling
Author(s) -
Berlew Erin E.,
Kuznetsov Ivan A.,
Yamada Keisuke,
Bugaj Lukasz J.,
Boerckel Joel D.,
Chow Brian Y.
Publication year - 2021
Publication title -
advanced biology
Language(s) - English
Resource type - Journals
ISSN - 2701-0198
DOI - 10.1002/adbi.202100810
Subject(s) - rhoa , microbiology and biotechnology , adherens junction , cytoskeleton , optogenetics , gtpase activating protein , small gtpase , biology , mechanotransduction , gtpase , signal transduction , chemistry , cell , g protein , neuroscience , cadherin , biochemistry
Optogenetic tools are created to control RhoA GTPase, a central regulator of actin organization and actomyosin contractility. RhoA GTPase, or its upstream activator ARHGEF11, is fused to BcLOV4, a photoreceptor that can be dynamically recruited to the plasma membrane by a light‐regulated protein‐lipid electrostatic interaction with the inner leaflet. Direct membrane recruitment of these proteins induces potent contractile signaling sufficient to separate adherens junctions with as little as one pulse of blue light. Induced cytoskeletal morphology changes are dependent on the alignment of the spatially patterned stimulation with the underlying cell polarization. RhoA‐mediated cytoskeletal activation drives yes‐associated protein (YAP) nuclear localization within minutes and consequent mechanotransduction verified by YAP‐transcriptional enhanced associate domain transcriptional activity. These single‐transgene tools do not require protein binding partners for dynamic membrane localization and permit spatiotemporally precise control over RhoA signaling to advance the study of its diverse regulatory roles in cell migration, morphogenesis, and cell cycle maintenance.