Detection of partially phosphorylated forms of ERK by monoclonal antibodies reveals spatial regulation of ERK activity by phosphatases.
When cells are stimulated by mitogens, extracellular signal-regulated kinase (ERK) is activated by phosphorylation of its regulatory threonine (Thr) and tyrosine (Tyr) residues. The inactivation of ERK may occur by phosphatase-mediated removal of the phosphates from these Tyr, Thr or both residues together. In this study, antibodies that selectively recognize all combinations of phosphorylation of the regulatory Thr and Tyr residues of ERK were developed, and used to study the inactivation of ERK upon mitogenic stimulation. We found that inactivation of ERK in the early stages of mitogenic stimulation involves separate Thr and Tyr phosphatases which operate differently in the nucleus and in the cytoplasm. Thus, ERK is differentially regulated in various subcellular compartments to secure proper length and strength of activation, which eventually determine the physiological outcome of many external signals.
Yao
Z
Z
Department of Biological Regulation, The Weizmann Institute of Science, Rehovot, Israel.
Dolginov
Y
Y
Hanoch
T
T
Yung
Y
Y
Ridner
G
G
Lando
Z
Z
Zharhary
D
D
Seger
R
R
D000818:Animals; D000911:Antibodies, Monoclonal / Q000378:metabolism*; D000918:Antibody Specificity / Q000276:immunology; D001665:Binding Sites / Q000276:immunology; D015153:Blotting, Western; D016466:CHO Cells; D002467:Cell Nucleus / Q000201:enzymology; D006224:Cricetinae; D003593:Cytoplasm / Q000201:enzymology; D004789:Enzyme Activation / Q000187:drug effects; D004797:Enzyme-Linked Immunosorbent Assay; D004815:Epidermal Growth Factor / Q000494:pharmacology; D066246:ErbB Receptors / Q000235:genetics; D007424:Intracellular Fluid / Q000201:enzymology; D019950:Mitogen-Activated Protein Kinase 1 / Q000037:antagonists & inhibitors / Q000276:immunology / Q000378:metabolism*; D048052:Mitogen-Activated Protein Kinase 3; D020928:Mitogen-Activated Protein Kinases / Q000037:antagonists & inhibitors / Q000276:immunology / Q000378:metabolism*; D008934:Mitogens / Q000494:pharmacology; D010744:Phosphoric Monoester Hydrolases / Q000378:metabolism* / Q000494:pharmacology; D010766:Phosphorylation; D015398:Signal Transduction / Q000187:drug effects; D013912:Threonine / Q000276:immunology / Q000378:metabolism; D014162:Transfection; D014443:Tyrosine / Q000276:immunology / Q000378:metabolism
D000911:Antibodies, Monoclonal; D008934:Mitogens; D013912:Threonine; D014443:Tyrosine; D004815:Epidermal Growth Factor; D066246:ErbB Receptors; D019950:Mitogen-Activated Protein Kinase 1; D048052:Mitogen-Activated Protein Kinase 3; D020928:Mitogen-Activated Protein Kinases; D010744:Phosphoric Monoester Hydrolases