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Automated FRET quantification shows distinct subcellular ERK activation kinetics in response to graded EGFR signaling in Drosophila
Author(s) -
Ogura Yosuke,
Sami Mustafa M.,
Wada Housei,
Hayashi Shigeo
Publication year - 2019
Publication title -
genes to cells
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.912
H-Index - 115
eISSN - 1365-2443
pISSN - 1356-9597
DOI - 10.1111/gtc.12679
Subject(s) - mapk/erk pathway , förster resonance energy transfer , biology , microbiology and biotechnology , signal transduction , receptor tyrosine kinase , kinase , cytoplasm , subcellular localization , tyrosine kinase , fluorescence , physics , quantum mechanics
Threshold responses to an activity gradient allow a single signaling pathway to yield multiple outcomes. Extracellular signal‐regulated kinase (ERK) is one such signal, which couples receptor tyrosine kinase signaling with multiple cellular responses in various developmental processes. Recent advances in the development of fluorescent biosensors for live imaging have enabled the signaling activities accompanying embryonic development to be monitored in real time. Here, we used an automated computational program to quantify the signals of a fluorescence resonance energy transfer (FRET) reporter for activated ERK, and we used this system to monitor the spatio‐temporal dynamics of ERK during neuroectoderm patterning in Drosophila embryos. We found that the cytoplasmic and nuclear ERK activity gradients show distinct kinetics in response to epidermal growth factor receptor activation. The ERK activation patterns implied that the cytoplasmic ERK activity is modulated into a threshold response in the nucleus.