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Dev Cell. 2017 Sep 11;42(5):542-553.e4. doi: 10.1016/j.devcel.2017.07.014. Epub 2017 Aug 17.

A Real-Time Biosensor for ERK Activity Reveals Signaling Dynamics during C. elegans Cell Fate Specification.

Author information

1
Department of Biological Sciences, Columbia University, New York, NY, USA; Department of Biochemistry & Molecular Biophysics, Columbia University Medical Center, New York, NY, USA.
2
Department of Biological Sciences, Columbia University, New York, NY, USA.
3
Department of Molecular Biology & Genetics, The Johns Hopkins University School of Medicine, Baltimore, MD, USA. Electronic address: sregot@jhmi.edu.
4
Department of Biological Sciences, Columbia University, New York, NY, USA; Department of Biochemistry & Molecular Biophysics, Columbia University Medical Center, New York, NY, USA. Electronic address: isg4@columbia.edu.

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.

KEYWORDS:

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

PMID:
28826819
PMCID:
PMC5595649
DOI:
10.1016/j.devcel.2017.07.014
[Indexed for MEDLINE]
Free PMC Article

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