Abstract
Current understanding of cell regulatory systems suggests a diverse array of extracellular stimuli commonly recruit a limited cadre of core signal transduction modules to drive discrete stimulus-specific responses. One such module is the Raf-MEK-extracellular signal-regulated kinase (ERK) kinase cascade. Little information exists about how this pathway can be appropriately coupled to discrete cell biological processes. Contributing factors may include regulation of the duration, amplitude, and/or subcellular compartmentalization of active ERK1/2. To define properties of ERK1/2 that may help mediate stimulus-selective signal propagation, we have examined the dynamic behavior of native ERK1/2 activation at the single-cell level. In primary human cell cultures, ERK1/2 activation is not an all-or-none response. Instead, the amount of active ERK1/2 in individual cells accumulated in proportion to the concentration of external stimulus. The variable degree of ERK1/2 activation correlated well with the degree of ERK1/2 effector activation. Therefore, the relative amplitude of ERK1/2 activation within a cell can be modulated and may contribute to the generation of stimulus-specific biological responses. Importantly, we also found that the capacity of active ERK1/2 to accumulate in the nucleus and drive immediate-early gene expression is dependent upon the nature of the inductive signal, but independent of the amplitude of ERK1/2 activation. Therefore, nuclear accumulation of active ERK1/2 is a discrete regulated step that can direct the function of the kinase in response to specific stimuli.
MeSH Terms
Active Transport, Cell Nucleus
Blotting, Western
Cell Nucleus/enzymology,metabolism
Dose-Response Relationship, Drug
Enzyme Activation
Epidermal Growth Factor/metabolism
HeLa Cells
Humans
Immunoblotting
Ligands
Microscopy, Fluorescence
Mitogen-Activated Protein Kinase 1/metabolism
Mitogen-Activated Protein Kinase 3/metabolism
Mitogen-Activated Protein Kinases/metabolism
Phenotype
Proto-Oncogene Proteins c-fos/metabolism
Signal Transduction
Substrate Specificity
Tetradecanoylphorbol Acetate
Chemicals
Ligands
Proto-Oncogene Proteins c-fos
Epidermal Growth Factor
Mitogen-Activated Protein Kinase 1
Mitogen-Activated Protein Kinase 3
Mitogen-Activated Protein Kinases
Tetradecanoylphorbol Acetate
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Whitehurst Angelique
Department of Cell Biology, UT Southwestern Medical Center, 5323 Harry Hines Blvd., Dallas, TX 75390, USA.
Cobb Melanie H
White Michael A
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