RRC ID 65035
著者 Inaba H, Miao Q, Nakata T.
タイトル Optogenetic control of small GTPases reveals RhoA mediates intracellular calcium signaling.
ジャーナル J Biol Chem
Abstract Rho/Ras family small GTPases are known to regulate numerous cellular processes, including cytoskeletal reorganization, cell proliferation, and cell differentiation. These processes are also controlled by Ca2+, and consequently, crosstalk between these signals is considered likely. However, systematic quantitative evaluation has not yet been reported. To fill this gap, we constructed optogenetic tools to control the activity of small GTPases (RhoA, Rac1, Cdc42, Ras, Rap, and Ral) using an improved light-inducible dimer system (iLID). We characterized these optogenetic tools with genetically encoded red fluorescence intensity-based small GTPase biosensors and confirmed these optogenetic tools' specificities. Using these optogenetic tools, we investigated calcium mobilization immediately after small GTPase activation. Unexpectedly, we found that a transient intracellular calcium elevation was specifically induced by RhoA activation in RPE1 and HeLa cells. RhoA activation also induced transient intracellular calcium elevation in MDCK and HEK293T cells, suggesting that generally RhoA induces calcium signaling. Interestingly, the molecular mechanisms linking RhoA activation to calcium increases were shown to be different among the different cell types: In RPE1 and HeLa cells, RhoA activated phospholipase C epsilon (PLCε) at the plasma membrane, which in turn induced Ca2+ release from the endoplasmic reticulum (ER). The RhoA-PLCε axis induced calcium-dependent NFAT nuclear translocation, suggesting it does activate intracellular calcium signaling. Conversely, in MDCK and HEK293T cells, RhoA-ROCK-myosin II axis induced the calcium transients. These data suggest universal coordination of RhoA and calcium signaling in cellular processes, such as cellular contraction and gene expression.
巻・号 296
ページ 100290
公開日 2021-1-1
DOI 10.1016/j.jbc.2021.100290
PII S0021-9258(21)00058-2
PMID 33453281
PMC PMC7949103
MeSH Animals Biosensing Techniques / methods Calcium / metabolism* Calcium Signaling / genetics* Cell Differentiation Cell Proliferation Dogs Gene Expression Regulation HEK293 Cells HeLa Cells Humans Light Light Signal Transduction / genetics* Madin Darby Canine Kidney Cells Optogenetics / methods* Organ Specificity Phosphoinositide Phospholipase C / genetics Phosphoinositide Phospholipase C / metabolism cdc42 GTP-Binding Protein / genetics cdc42 GTP-Binding Protein / metabolism rac1 GTP-Binding Protein / genetics rac1 GTP-Binding Protein / metabolism ral GTP-Binding Proteins / genetics ral GTP-Binding Proteins / metabolism rap GTP-Binding Proteins / genetics rap GTP-Binding Proteins / metabolism ras Proteins / genetics ras Proteins / metabolism rhoA GTP-Binding Protein / genetics* rhoA GTP-Binding Protein / metabolism
IF 4.238
リソース情報
ヒト・動物細胞 HeLa(RCB0007)