Data-driven modeling reconciles kinetics of ERK phosphorylation, localization, and activity states.
The extracellular signal-regulated kinase (ERK) signaling pathway controls cell proliferation and differentiation in metazoans. Two hallmarks of its dynamics are adaptation of ERK phosphorylation, which has been linked to negative feedback, and nucleocytoplasmic shuttling, which allows active ERK to phosphorylate protein substrates in the nucleus and cytosol. To integrate these complex features, we acquired quantitative biochemical and live-cell microscopy data to reconcile phosphorylation, localization, and activity states of ERK. While maximal growth factor stimulation elicits transient ERK phosphorylation and nuclear translocation responses, ERK activities available to phosphorylate substrates in the cytosol and nuclei show relatively little or no adaptation. Free ERK activity in the nucleus temporally lags the peak in nuclear translocation, indicating a slow process. Additional experiments, guided by kinetic modeling, show that this process is consistent with ERK's modification of and release from nuclear substrate anchors. Thus, adaptation of whole-cell ERK phosphorylation is a by-product of transient protection from phosphatases. Consistent with this interpretation, predictions concerning the dose-dependence of the pathway response and its interruption by inhibition of MEK were experimentally confirmed.
Department of Chemical and Biomolecular Engineering, North Carolina State University, Raleigh, NC, USA.
Shoeb
S
Ahmed
Kyle G
KG
Grant
Laura E
LE
Edwards
Anisur
A
Rahman
Murat
M
Cirit
Michael B
MB
Goshe
Jason M
JM
Haugh
D021581:Active Transport, Cell Nucleus; D000818:Animals; D048049:Extracellular Signal-Regulated MAP Kinases / Q000737:chemistry* / Q000378:metabolism; D025461:Feedback, Physiological; D007700:Kinetics; D051379:Mice; D008962:Models, Theoretical; D041681:NIH 3T3 Cells; D010766:Phosphorylation; D021381:Protein Transport / Q000235:genetics; D017978:Receptors, Growth Factor / Q000235:genetics; D015398:Signal Transduction / Q000235:genetics
D017978:Receptors, Growth Factor; D048049:Extracellular Signal-Regulated MAP Kinases