A Live-Cell Screen for Altered Erk Dynamics Reveals Principles of Proliferative Control. Author Alexander Goglia, Maxwell Wilson, Siddhartha Jena, Jillian Silbert, Lena Basta, Danelle Devenport, Jared Toettcher Publication Year 2020 Type Journal Article Abstract 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. Keywords Animals, Mice, Signal Transduction, Cell Proliferation, High-Throughput Screening Assays, Phosphorylation, MAP Kinase Signaling System, ras Proteins, Optogenetics, Extracellular Signal-Regulated MAP Kinases, Keratinocytes, ErbB Receptors, Proto-Oncogene Proteins c-akt, Drug Evaluation, Preclinical Journal Cell Syst Volume 10 Issue 3 Pages 240-253.e6 Date Published 2020 Mar 25 ISSN Number 2405-4720 DOI 10.1016/j.cels.2020.02.005 Alternate Journal Cell Syst PMCID PMC7540725 PMID 32191874 PubMedPubMed CentralGoogle ScholarBibTeXEndNote X3 XML