@article{3a4d30fea0d34ab488392213b2503068,
title = "A Real-Time Biosensor for ERK Activity Reveals Signaling Dynamics during C. elegans Cell Fate Specification",
abstract = "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. A genetically encoded biosensor for ERK activity allows qualitative or quantitative assessment by analysis of individual nuclei during development and neural function in diverse cell contexts in C. elegans. In-depth analysis of ERK activity over time in the vulval precursor cells has identified dynamic features including pulsatile frequency-modulated signaling.",
keywords = "C. elegans, ERK, biosensor, extracellular signal-regulated kinase, kinase translocation reporter, vulval development",
author = "{de la Cova}, Claire and Robert Townley and Sergi Regot and Iva Greenwald",
note = "Funding Information: We gratefully acknowledge Markus Covert, Jake Hughey, and Sajia Akhter for assisting with custom software development; Ari Pani, Bob Goldstein, and David Matus for generously sharing reagents and advice for CRISPR/Cas9-mediated single-copy transgene insertion; Julie Canman for the gift of a VPC marker; and Swathi Arur and Tim Schedl for helpful discussion about the germline. We are also indebted to Daniel Shaye and Claudia Tenen for helpful comments on the manuscript. Some of the strains used in this study were provided by the Caenorhabditis Genetics Center, which is supported by the NIH Office of Research Infrastructure Programs ( P40 OD010440 ). This work was supported by Senior Scholar Award AG-SS-2951-12 from the Ellison Medical Foundation and grant R01 GM114140 from the NIH (to I.G.), as well as a 2016 Kimmel Scholar Award (to S.R.). Funding Information: We gratefully acknowledge Markus Covert, Jake Hughey, and Sajia Akhter for assisting with custom software development; Ari Pani, Bob Goldstein, and David Matus for generously sharing reagents and advice for CRISPR/Cas9-mediated single-copy transgene insertion; Julie Canman for the gift of a VPC marker; and Swathi Arur and Tim Schedl for helpful discussion about the germline. We are also indebted to Daniel Shaye and Claudia Tenen for helpful comments on the manuscript. Some of the strains used in this study were provided by the Caenorhabditis Genetics Center, which is supported by the NIH Office of Research Infrastructure Programs (P40 OD010440). This work was supported by Senior Scholar Award AG-SS-2951-12 from the Ellison Medical Foundation and grant R01 GM114140 from the NIH (to I.G.), as well as a 2016 Kimmel Scholar Award (to S.R.). Publisher Copyright: {\textcopyright} 2017",
year = "2017",
month = sep,
day = "11",
doi = "10.1016/j.devcel.2017.07.014",
language = "English (US)",
volume = "42",
pages = "542--553.e4",
journal = "Developmental Cell",
issn = "1534-5807",
publisher = "Cell Press",
number = "5",
}