Kinetics of receptor tyrosine kinase activation define ERK signaling dynamics

Abstract

In responses to activation of receptor tyrosine kinases (RTKs), crucial cell fate decisions depend on the duration and dynamics of ERK signaling. In PC12 cells, epidermal growth factor (EGF) induces transient ERK activation that leads to cell proliferation, whereas nerve growth factor (NGF) promotes sustained ERK activation and cell differentiation. These differences have typically been assumed to reflect distinct feedback mechanisms in the Raf-MEK-ERK signaling network, with the receptors themselves acting as simple upstream inputs. We failed to confirm the expected differences in feedback type when investigating transient versus sustained signaling downstream of the EGF receptor (EGFR) and NGF receptor (TrkA). Instead, we found that ERK signaling faithfully followed RTK dynamics when receptor signaling was modulated in different ways. EGFR activation kinetics, and consequently ERK signaling dynamics, were switched from transient to sustained when receptor internalization was inhibited with drugs or mutations, or when cells expressed a chimeric receptor likely to have impaired dimerization. In addition, EGFR and ERK signaling both became more sustained when substoichiometric levels of erlotinib were added to reduce duration of EGFR kinase activation. Our results argue that RTK activation kinetics play a crucial role in determining MAP kinase cascade signaling dynamics and cell fate decisions, and that signaling outcome can be modified by activating a given RTK in different ways.

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BoxCarmax: A High-Selectivity Data-Independent Acquisition Mass Spectrometry Method for the Analysis of Protein Turnover and Complex Samples

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A Circuit for Secretion-coupled Cellular Autonomy in Multicellular Eukaryotes

“…First, it is noteworthy that both PM-proximal events as well as the endomembrane-localized events peak simultaneously at ~5 min post EGF stimulation, e.g., EGF-triggered pERK (47,48), coupling of ligand-activated EGFR to trimeric Gi at the PM (49), and Golgi localized events such as Arf1 activation (Figure 4A). Because cells frequently respond with high fidelity and specificity in an ultrasensitive fashion to graded input (50), such that a response is triggered only when signals reach peak levels and break thresholds, our findings imply that despite a propagation delay, there is virtually no latency between when these peak signals are reached.…”