Details

Autor(en) / Beteiligte

• Goglia, Alexander G.
• Wilson, Maxwell Z.
• Jena, Siddhartha G.
• Silbert, Jillian
• Basta, Lena P.
• Devenport, Danelle
• Toettcher, Jared E.

Titel

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

Ist Teil von

• Cell systems, 2020-03, Vol.10 (3), p.240-253.e6

Ort / Verlag

United States: Elsevier Inc

Erscheinungsjahr

2020

Link zum Volltext

Quelle

Elektronische Zeitschriftenbibliothek - Frei zugängliche E-Journals

Beschreibungen/Notizen

• 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. [Display omitted] •Keratinocytes exhibit robust, rapid Erk pulses even without exogenous growth factors•Screening 429 kinase inhibitors reveals distinct perturbations to Erk dynamics•Receptor tyrosine kinase inhibition can increase Erk activity and pulse frequency•Optogenetic control reveals how Erk dynamics alter cell proliferation Goglia et al. identified modulators of Erk dynamics by screening a library of 429 kinase inhibitors and monitoring Erk activity over 5 h in more than 80,000 single primary mouse keratinocytes. They identified both known and uncharacterized modulators, including inhibitors of non-EGFR receptor tyrosine kinases (RTKs) that increased Erk pulse frequency and overall activity. Their 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.