title: Epidermal growth factor activation of vinculin and beta 1-integrin gene transcription in quiescent Swiss 3T3 cells. Regulation through a protein kinase C-independent pathway.
abstract: Growth activation of quiescent Swiss 3T3 fibroblasts leads to a rapid induction of vinculin and beta 1-integrin gene expression. Addition of serum, epidermal growth factor (EGF), or platelet-derived growth factor to serum-starved, density-arrested cells resulted in a rapid increase in vinculin and beta 1-integrin mRNA levels and a corresponding increase in vinculin synthesis. The increase in vinculin and beta 1-integrin mRNA expression by serum or EGF was not blocked by the inhibition of protein synthesis by cycloheximide. The kinetics of induction of vinculin and beta 1-integrin mRNAs by EGF are different: vinculin mRNA levels reached a peak of expression 4-5-fold greater than that measured in quiescent cells by 2 h after addition of growth factor, whereas beta 1-integrin mRNA levels increased more slowly and to a lesser extent, reaching peaks of 2-3-fold induction at 5 h poststimulation. Down-regulation of protein kinase C by prolonged pretreatment of cells with phorbol 1,2-myristate 1,3-acetate had no effect on the ability of EGF or platelet-derived growth factor to activate vinculin or beta 1-integrin mRNA expression. Furthermore, direct activation of protein kinase C with 1,2-myristate 1,3-acetate did not induce the expression of vinculin or beta 1-integrin mRNA, but did activate c-fos expression. In vitro nuclear "run-on" transcription assays demonstrate a greater than 7-fold increase in vinculin and beta 1-integrin transcription at 40-60 min after addition of EGF when compared with levels in quiescent cells. This activation was rapid and transient, but appeared to occur later than the increase in c-fos and actin transcription. These results demonstrate that vinculin and beta 1-integrin, important components of the cell adhesion apparatus, are members of a group of immediate early growth-responsive genes, along with c-fos, c-myc, actin, and fibronectin. In addition, regulation of these cell adhesion genes occurs exclusively through a protein kinase C-independent pathway in serum-deprived, density-arrested Swiss 3T3 cells.
authors.affiliation: Department of Biochemistry, Boston University School of Medicine, Massachusetts.
authors.forename: R E
authors.initials: RE
authors.lastname: Bellas
authors.forename: R
authors.initials: R
authors.lastname: Bendori
authors.forename: S R
authors.initials: SR
authors.lastname: Farmer
mesh_terms: D000818:Animals; D015152:Blotting, Northern; D015153:Blotting, Western; D003598:Cytoskeletal Proteins / Q000235:genetics* / Q000378:metabolism; D015180:Electrophoresis, Gel, Two-Dimensional; D004789:Enzyme Activation; D004815:Epidermal Growth Factor / Q000494:pharmacology*; D005347:Fibroblasts / Q000378:metabolism; D016023:Integrins / Q000235:genetics* / Q000378:metabolism; D051379:Mice; D011493:Protein Kinase C / Q000378:metabolism; D012313:RNA / Q000302:isolation & purification; D012333:RNA, Messenger / Q000032:analysis; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D014158:Transcription, Genetic; D016596:Vinculin
chemical_list: D003598:Cytoskeletal Proteins; D016023:Integrins; D012333:RNA, Messenger; D016596:Vinculin; D004815:Epidermal Growth Factor; D012313:RNA; D011493:Protein Kinase C; D013755:Tetradecanoylphorbol Acetate