Developmental Cell
Volume 42, Issue 5, 11 September 2017, Pages 542-553.e4
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A Real-Time Biosensor for ERK Activity Reveals Signaling Dynamics during C. elegans Cell Fate Specification

https://doi.org/10.1016/j.devcel.2017.07.014Get rights and content
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Highlights

  • ERK-nKTR is a genetically encoded biosensor for extracellular signal-regulated kinase

  • Qualitative or quantitative assessment is possible by analyzing individual nuclei

  • ERK-nKTR is a faithful reporter for ERK activity in several different cell types

  • In the vulval precursor cells, there is pulsatile, frequency-modulated signaling

Summary

Kinase translocation reporters (KTRs) are genetically encoded fluorescent activity sensors that convert kinase activity into a nucleocytoplasmic shuttling equilibrium for visualizing single-cell signaling dynamics. Here, we adapt the first-generation KTR for extracellular signal-regulated kinase (ERK) to allow easy implementation in vivo. This sensor, “ERK-nKTR,” allows quantitative and qualitative assessment of ERK activity by analysis of individual nuclei and faithfully reports ERK activity during development and neural function in diverse cell contexts in Caenorhabditis elegans. Analysis of ERK activity over time in the vulval precursor cells, a well-characterized paradigm of epidermal growth factor receptor (EGFR)-Ras-ERK signaling, has identified dynamic features not evident from analysis of developmental endpoints alone, including pulsatile frequency-modulated signaling associated with proximity to the EGF source. The toolkit described here will facilitate studies of ERK signaling in other C. elegans contexts, and the design features will enable implementation of this technology in other multicellular organisms.

Keywords

extracellular signal-regulated kinase
ERK
biosensor
kinase translocation reporter
C. elegans
vulval development

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