ArticleBMC biology2015
Dynamic single cell measurements of kinase activity by synthetic kinase activity relocation sensors.
Article in BMC biology, 2015. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 19 papers.
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Who cites it
19 citing papers in PubMed, 38 citations in OpenAlex.
- Noninvasive methods to monitor dynamic single-cell events.Trends in cell biology · 2026Review
- Tools for live-cell imaging of cytoskeletal and nuclear behavior in the unconventional yeast,Molecular biology of the cell · 2024Article
- Positive feedback induces switch between distributive and processive phosphorylation of Hog1.Nature communications · 2023Article
- Exploratory polarization facilitates mating partner selection inMolecular biology of the cell · 2021Article
- Article
- Expanding the Toolkit of Fluorescent Biosensors for Studying Mitogen Activated Protein Kinases in Plants.International journal of molecular sciences · 2020Article
- Ratiometric GPCR signaling enables directional sensing in yeast.PLoS biology · 2019Article
- Mechanical stress impairs pheromone signaling via Pkc1-mediated regulation of the MAPK scaffold Ste5.The Journal of cell biology · 2019Article
- Phosphorylation-dependent protein design: design of a minimal protein kinase-inducible domain.Organic & biomolecular chemistry · 2019Article
- MAPK modulation of yeast pheromone signaling output and the role of phosphorylation sites in the scaffold protein Ste5.Molecular biology of the cell · 2019Article
- Genetically Encoded Fluorescent Biosensors Illuminate the Spatiotemporal Regulation of Signaling Networks.Chemical reviews · 2018Review
- Timing of gene expression in a cell-fate decision system.Molecular systems biology · 2018Article
- CDK and MAPK Synergistically Regulate Signaling Dynamics via a Shared Multi-site Phosphorylation Region on the Scaffold Protein Ste5.Molecular cell · 2018Article
- Protein kinase C and calcineurin cooperatively mediate cell survival under compressive mechanical stress.Proceedings of the National Academy of Sciences of the United States of America · 2017Article
- Mitogen-activated protein kinase (MAPK) dynamics determine cell fate in the yeast mating response.The Journal of biological chemistry · 2017Article
- Nuclear relocation of Kss1 contributes to the specificity of the mating response.Scientific reports · 2017Article
- Single-cell dynamics and variability of MAPK activity in a yeast differentiation pathway.Proceedings of the National Academy of Sciences of the United States of America · 2016Article
- Rewiring MAP kinases in Saccharomyces cerevisiae to regulate novel targets through ubiquitination.eLife · 2016Article
- Real-time quantification of protein expression at the single-cell level via dynamic protein synthesis translocation reporters.Nature communications · 2016Article
Corrections and comments
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Authors and funding
3 authors at 1 institution in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
backgroundMitogen activated protein kinases (MAPK) play an essential role in integrating extra-cellular signals and intra-cellular cues to allow cells to grow, adapt to stresses, or undergo apoptosis. Budding yeast serves as a powerful system to understand the fundamental regulatory mechanisms that allow these pathways to combine multiple signals and deliver an appropriate response. To fully comprehend the variability and dynamics of these signaling cascades, dynamic and quantitative single cell measurements are required. Microscopy is an ideal technique to obtain these data; however, novel assays have to be developed to measure the activity of these cascades.
resultsWe have generated fluorescent biosensors that allow the real-time measurement of kinase activity at the single cell level. Here, synthetic MAPK substrates were engineered to undergo nuclear-to-cytoplasmic relocation upon phosphorylation of a nuclear localization sequence. Combination of fluorescence microscopy and automated image analysis allows the quantification of the dynamics of kinase activity in hundreds of single cells. A large heterogeneity in the dynamics of MAPK activity between individual cells was measured. The variability in the mating pathway can be accounted for by differences in cell cycle stage, while, in the cell wall integrity pathway, the response to cell wall stress is independent of cell cycle stage.
conclusionsThese synthetic kinase activity relocation sensors allow the quantification of kinase activity in live single cells. The modularity of the architecture of these reporters will allow their application in many other signaling cascades. These measurements will allow to uncover new dynamic behaviour that previously could not be observed in population level measurements.
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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.