A Live-Cell Screen for Altered Erk Dynamics Reveals Principles of Proliferative Control.

TitleA Live-Cell Screen for Altered Erk Dynamics Reveals Principles of Proliferative Control.
Publication TypeJournal Article
Year of Publication2020
AuthorsGoglia, AG, Wilson, MZ, Jena, SG, Silbert, J, Basta, LP, Devenport, D, Toettcher, JE
JournalCell Syst
Date Published2020 03 25
KeywordsAnimals, Cell Proliferation, Drug Evaluation, Preclinical, ErbB Receptors, Extracellular Signal-Regulated MAP Kinases, High-Throughput Screening Assays, Keratinocytes, MAP Kinase Signaling System, Mice, Optogenetics, Phosphorylation, Proto-Oncogene Proteins c-akt, ras Proteins, Signal Transduction

<p>Complex, time-varying responses have been observed widely in cell signaling, but how specific dynamics are generated or regulated is largely unknown. One major obstacle has been that high-throughput screens are typically incompatible with the live-cell assays used to monitor dynamics. Here, we address this challenge by screening a library of 429 kinase inhibitors and monitoring extracellular-regulated kinase (Erk) activity over 5 h in more than 80,000 single primary mouse keratinocytes. Our screen reveals both known and uncharacterized modulators of Erk dynamics, including inhibitors of non-epidermal growth factor receptor (EGFR) receptor tyrosine kinases (RTKs) that increase Erk pulse frequency and overall activity. Using drug treatment and direct optogenetic control, we demonstrate that drug-induced changes to Erk dynamics alter the conditions under which cells proliferate. Our work opens the door to high-throughput screens using live-cell biosensors and reveals that cell proliferation integrates information from Erk dynamics as well as additional permissive cues.</p>

Alternate JournalCell Syst
PubMed ID32191874
PubMed Central IDPMC7540725
Grant ListDP2 EB024247 / EB / NIBIB NIH HHS / United States
F30 CA206408 / CA / NCI NIH HHS / United States
T32 GM007388 / GM / NIGMS NIH HHS / United States