Intensiometric biosensors visualize the activity of multiple small GTPases in vivo

Kim, Jihoon; Lee, Sangkyu; Jung, Kanghoon; Oh, Won Chan; Kim, Nury; Son, Seungkyu; Jo, YoungJu; Kwon, Hyung Bae; Heo, Won Do

doi:10.1038/s41467-018-08217-3

Cited by 31 publications

(31 citation statements)

References 37 publications

(63 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Although relatively recently established, KTRs offer several potential advantages in comparison to FRET biosensors, such as a wider dynamic range and seemingly more accurate reporting of the downregulation of kinase activity [ 51 , 53 ]. Although KTRs have been successfully implemented in mammalian cells, Caenorhabditis elegans, and zebrafish [ 51 , 52 , 53 , 54 , 55 ], their use has not yet been reported in plants. In order to expand the set of fluorescent reporters available for studying MAPK activation in plants, we were therefore interested in determining if the KTR principle could be applied to Arabidopsis .…”

Section: Resultsmentioning

confidence: 99%

“…By attaching a docking domain targeted by a kinase of interest to this type of biosensor, the nucleocytoplasmic shuttling of the sensor can serve as a visual indicator of kinase activity [ 51 ]. Translocation biosensors offer several potential benefits in comparison to FRET-based biosensors and have proven to be powerful tools for documenting kinase spatiotemporal dynamics in different animal models [ 51 , 52 , 53 , 54 , 55 ].…”

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Expanding the Toolkit of Fluorescent Biosensors for Studying Mitogen Activated Protein Kinases in Plants

Seitz

Krysan

2020

IJMS

View full text Add to dashboard Cite

Mitogen-activated protein kinases (MAPKs) are key regulators of numerous biological processes in plants. To better understand the mechanisms by which these kinases function, high resolution measurement of MAPK activation kinetics in different biological contexts would be beneficial. One method to measure MAPK activation in plants is via fluorescence-based genetically-encoded biosensors, which can provide real-time readouts of the temporal and spatial dynamics of kinase activation in living tissue. Although fluorescent biosensors have been widely used to study MAPK dynamics in animal cells, there is currently only one MAPK biosensor that has been described for use in plants. To facilitate creation of additional plant-specific MAPK fluorescent biosensors, we report the development of two new tools: an in vitro assay for efficiently characterizing MAPK docking domains and a translocation-based kinase biosensor for use in plants. The implementation of these two methods has allowed us to expand the available pool of plant MAPK biosensors, while also providing a means to generate more specific and selective MAPK biosensors in the future. Biosensors developed using these methods have the potential to enhance our understanding of the roles MAPKs play in diverse plant signaling networks affecting growth, development, and stress response.

show abstract

Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Expanding the Toolkit of Fluorescent Biosensors for Studying Mitogen Activated Protein Kinases in Plants

Seitz

Krysan

2020

IJMS

View full text Add to dashboard Cite

show abstract

“…More recently, the ddFP-based biosensors for small GTPases have been introduced [ 55 ]. These intermolecular biosensors are composed of two parts; the first part is the GTPase itself attached to GA, and the second part is the copy-B-tagged the RBD domain from its corresponding effector.…”

Section: Sensing Strategies Of Fp-based Biosensorsmentioning

confidence: 99%

“…When the activated KRas binds to the RBD domain from its effector Raf1, the heterodimer between GA and copy-B can be formed to increase the brightness of green fluorescence, representing the active state of KRas in live cells. These ddFP-based biosensors for small GTPases were successfully utilized to monitor the spatiotemporal activity of the small GTPases in the single dendritic spines and brains of freely behaving mice [ 55 ].…”

Section: Sensing Strategies Of Fp-based Biosensorsmentioning

confidence: 99%

Genetically Encoded Biosensors Based on Fluorescent Proteins

Kim

Lee

et al. 2021

Sensors

View full text Add to dashboard Cite

Genetically encoded biosensors based on fluorescent proteins (FPs) allow for the real-time monitoring of molecular dynamics in space and time, which are crucial for the proper functioning and regulation of complex cellular processes. Depending on the types of molecular events to be monitored, different sensing strategies need to be applied for the best design of FP-based biosensors. Here, we review genetically encoded biosensors based on FPs with various sensing strategies, for example, translocation, fluorescence resonance energy transfer (FRET), reconstitution of split FP, pH sensitivity, maturation speed, and so on. We introduce general principles of each sensing strategy and discuss critical factors to be considered if available, then provide representative examples of these FP-based biosensors. These will help in designing the best sensing strategy for the successful development of new genetically encoded biosensors based on FPs.

show abstract

“…The most successful strategies employed to date use Förster resonance energy transfer (FRET)-based sensors (e.g., see [176,177]), which have been highly informative and sensitive in cell-culture systems. While their translation to transgenic models for in vivo use have not been as successful, several recent examples using FRET, as well as other approaches, including measuring nuclear-to-cytoplasmic ratio changes in fluorescent sensors, have shown promise [178,179].…”

Section: Perspectivesmentioning

confidence: 99%

Receptor Tyrosine Kinases in Development: Insights from Drosophila

Mele

Johnson

2019

IJMS

View full text Add to dashboard Cite

Cell-to-cell communication mediates a plethora of cellular decisions and behaviors that are crucial for the correct and robust development of multicellular organisms. Many of these signals are encoded in secreted hormones or growth factors that bind to and activate cell surface receptors, to transmit the cue intracellularly. One of the major superfamilies of cell surface receptors are the receptor tyrosine kinases (RTKs). For nearly half a century RTKs have been the focus of intensive study due to their ability to alter fundamental aspects of cell biology, such as cell proliferation, growth, and shape, and because of their central importance in diseases such as cancer. Studies in model organisms such a Drosophila melanogaster have proved invaluable for identifying new conserved RTK pathway components, delineating their contributions, and for the discovery of conserved mechanisms that control RTK-signaling events. Here we provide a brief overview of the RTK superfamily and the general mechanisms used in their regulation. We further highlight the functions of several RTKs that govern distinct cell-fate decisions in Drosophila and explore how their activities are developmentally controlled.

show abstract

Intensiometric biosensors visualize the activity of multiple small GTPases in vivo

Cited by 31 publications

References 37 publications

Expanding the Toolkit of Fluorescent Biosensors for Studying Mitogen Activated Protein Kinases in Plants

Expanding the Toolkit of Fluorescent Biosensors for Studying Mitogen Activated Protein Kinases in Plants

Genetically Encoded Biosensors Based on Fluorescent Proteins

Receptor Tyrosine Kinases in Development: Insights from Drosophila

Contact Info

Product

Resources

About