ArticleJournal of immunology (Baltimore, Md. : 1950)2016
Optical Tools To Study the Isoform-Specific Roles of Small GTPases in Immune Cells.
Article in Journal of immunology (Baltimore, Md. : 1950), 2016. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 20 papers.
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Who cites it
20 citing papers in PubMed, 25 citations in OpenAlex.
- Article
- A new single chain, genetically encoded biosensor for RhoB GTPase based on FRET, useful for live-cell imaging.bioRxiv : the preprint server for biology · 2026Article
- Fluorescence Lifetime Imaging Application to Probe GTPase Activation in Macrophage Cell Line, Using the Time-Domain FastFLIM Modality.Methods in molecular biology (Clifton, N.J.) · 2026Article
- Unravelling molecular dynamics in living cells: Fluorescent protein biosensors for cell biology.Journal of microscopy · 2025Review
- Real-time imaging reveals a role for macrophage protrusive motility in melanoma invasion.The Journal of cell biology · 2025Article
- A proliferative to invasive switch is mediated by srGAP1 downregulation through the activation of TGF-β2 signaling.Cell reports · 2022Article
- Förster resonance energy transfer biosensors for fluorescence and time-gated luminescence analysis of rac1 activity.Scientific reports · 2022Article
- Förster resonance energy transfer biosensors for fluorescence and time-gated luminescence analysis of rac1 activity.Scientific reports · 2022Article
- Resource for FRET-Based Biosensor Optimization.Frontiers in cell and developmental biology · 2022Review
- Seeing is believing: tools to study the role of Rho GTPases during cytokinesis.Small GTPases · 2022Review
- TC10 regulates breast cancer invasion and metastasis by controlling membrane type-1 matrix metalloproteinase at invadopodia.Communications biology · 2021Article
- Metalloprotease ADAMTS-1 decreases cell migration and invasion modulating the spatiotemporal dynamics of Cdc42 activity.Cellular signalling · 2021Article
- Multiplex Imaging of Rho GTPase Activities in Living Cells.Methods in molecular biology (Clifton, N.J.) · 2021Article
- Optogenetics: Rho GTPases Activated by Light in Living Macrophages.Methods in molecular biology (Clifton, N.J.) · 2020Article
- Macrophage polarization impacts tunneling nanotube formation and intercellular organelle trafficking.Scientific reports · 2019Article
- Direct multiplex imaging and optogenetics of Rho GTPases enabled by near-infrared FRET.Nature chemical biology · 2018Article
- Characterization of Genetically Encoded FRET Biosensors for Rho-Family GTPases.Methods in molecular biology (Clifton, N.J.) · 2018Article
- Rac3 regulates breast cancer invasion and metastasis by controlling adhesion and matrix degradation.The Journal of cell biology · 2017Article
- The Role of Rho-GTPases and actin polymerization during Macrophage Tunneling Nanotube Biogenesis.Scientific reports · 2017Article
- Controllable Photodynamic Therapy Implemented by Regulating Singlet Oxygen Efficiency.Advanced science (Weinheim, Baden-Wurttemberg, Germany) · 2017Review
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Authors and funding
5 authors at 1 institution in 1 country.
Funding
Abstract
Despite the 92% homology of the hematopoietic cell-specific Rac2 to the canonical isoform Rac1, these isoforms have been shown to play nonredundant roles in immune cells. To study isoform-specific dynamics of Rac in live cells, we developed a genetically encoded, single-chain FRET-based biosensor for Rac2. We also made significant improvements to our existing single-chain Rac1 biosensor. We optimized the biosensor constructs for facile expression in hematopoietic cells and performed functional validations in murine macrophage sublines of RAW264.7 cells. Rac2, Rac1, and Cdc42 have been implicated in the formation of actin-rich protrusions by macrophages, but their individual activation dynamics have not been previously characterized. We found that both Rac1 and Rac2 had similar activation kinetics, yet they had distinct spatial distributions in response to the exogenous stimulus, fMLF. Active Rac1 was mainly localized to the cell periphery, whereas active Rac2 was distributed throughout the cell, with an apparent higher concentration in the perinuclear region. We also performed an extensive morphodynamic analysis of Rac1, Rac2, and Cdc42 activities during the extension of random protrusions. We found that Rac2 appears to play a leading role in the generation of random protrusions, as we observed an initial strong activation of Rac2 in regions distal from the leading edge, followed by the activation of Rac1, a second burst of Rac2 and then Cdc42 immediately behind the leading edge. Overall, isoform-specific biosensors that have been optimized for expression should be valuable for interrogating the coordination of Rho family GTPase activities in living cells.
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Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.