Actin polymerization is required for negative feedback regulation of epidermal growth factor-induced signal transduction.
Epidermal growth factor (EGF) induces rapid actin filament assembly in the membrane skeleton of a variety of cells. To investigate the significance of this process for signal transduction, actin polymerization is inhibited by dihydrocytochalasin B (CB). CB almost completely abolishes EGF-induced actin polymerization, as assessed by quantitative confocal laser scanning microscopy. Under these conditions, EGF induces enhanced EGF receptor (EGFR) tyrosine kinase activity, as well as superinduction of the c-fos proto-oncogene. These data suggest that EGF-induced actin polymerization may be important for negative feedback regulation of signal transduction by the EGFR. The phosphorylation of Thr654 by protein kinase C (PKC) is a well-characterized negative feedback control mechanism for signal transduction by the EGFR tyrosine kinase. A synthetic peptide, corresponding to the regions flanking Thr654 of the EGFR, is used to analyze EGF stimulated PKC activity by incorporation of 32P into the peptide. Cotreatment of cells with CB and EGF results in a complete loss of EGF-induced phosphorylation of the peptide. These data suggest that actin polymerization is obligatory for negative feedback regulation of the EGFR tyrosine kinase through the C-kinase pathway.
Rijken
P J
PJ
Department of Molecular Cell Biology, University of Utrecht, Padualaan 8, 3584 CH Utrecht, The Netherlands.
van Hal
G J
GJ
van der Heyden
M A
MA
Verkleij
A J
AJ
Boonstra
J
J
D000199:Actins / Q000378:metabolism*; D001704:Biopolymers; D002384:Catalysis; D003571:Cytochalasin B / Q000031:analogs & derivatives / Q000494:pharmacology; D004815:Epidermal Growth Factor / Q000378:metabolism* / Q000494:pharmacology; D066246:ErbB Receptors / Q000378:metabolism; D005786:Gene Expression Regulation; D006801:Humans; D009857:Oncogenes; D010766:Phosphorylation; D011493:Protein Kinase C / Q000378:metabolism; D000090063:Proto-Oncogene Mas; D016760:Proto-Oncogene Proteins c-fos / Q000235:genetics; D015398:Signal Transduction; D013912:Threonine / Q000378:metabolism; D014407:Tumor Cells, Cultured
D000199:Actins; D001704:Biopolymers; C000715470:MAS1 protein, human; D000090063:Proto-Oncogene Mas; D016760:Proto-Oncogene Proteins c-fos; D013912:Threonine; C024794:dihydrocytochalasin B; D003571:Cytochalasin B; D004815:Epidermal Growth Factor; D066246:ErbB Receptors; D011493:Protein Kinase C