Phorbol ester modulation of integrin-mediated cell adhesion: a postreceptor event.
Chinese hamster ovary (CHO) suspension culture cells adhere readily to substrata coated with extracellular matrix proteins such as fibronectin, vitronectin, or laminin. In the case of fibronectin, it is known that adhesion is mediated by an integrin-type, cell surface fibronectin receptor (FnR). We demonstrate here that treatment of CHO cells with submicromolar concentrations of phorbol ester produces a remarkable increase in the ability of these cells to adhere to fibronectin. Both the rate of adhesion and the efficiency of adhesion are enhanced about four- to fivefold. Further, phorbol ester treatment renders the fibronectin-mediated adhesion process less sensitive to inhibitors, including GRGDSP peptide and PB1, a monoclonal anti-FnR antibody. By contrast, nonspecific adhesion processes, for example cell attachment to substrata coated with polylysine or concanavalin A, are not affected by phorbol ester treatment. Thus integrin-mediated adhesion is modulated by phorbol esters, but nonspecific adhesion is not. Neither the number of cell surface FnRs nor the receptor affinity, as measured by 125I-fibronectin and 125I-anti-FnR antibody binding, is altered by phorbol ester treatment. Thus, the effect of phorbol ester on cell adhesion seems to occur at a step subsequent to initial ligand-receptor binding events. Since phorbol ester is a potent activator of protein kinase C, we examined phosphorylation patterns in control and phorbol-treated cells. In immunoprecipitates of lysates from suspension culture cells, there was no evidence of phorbol ester-stimulated phosphorylation of FnR or of talin, a protein thought to interact with FnR. These results suggest that phorbol ester effects on fibronectin-dependent adhesion are not due to phosphorylation of the FnR itself but rather may be due to postreceptor events, possibly the phosphorylation of cytoskeletal proteins involved in integrin-mediated adhesion.
Danilov
Y N
YN
Department of Pharmacology, School of Medicine, University of North Carolina, Chapel Hill 27599-7365.
Juliano
R L
RL
D000818:Animals; D002448:Cell Adhesion / Q000187:drug effects*; D002460:Cell Line; D003208:Concanavalin A / Q000494:pharmacology; D005353:Fibronectins; D016023:Integrins; D007700:Kinetics; D008562:Membrane Glycoproteins / Q000502:physiology*; D011107:Polylysine / Q000494:pharmacology; D011971:Receptors, Immunologic / Q000187:drug effects / Q000502:physiology*; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology*
D005353:Fibronectins; D016023:Integrins; D008562:Membrane Glycoproteins; D011971:Receptors, Immunologic; D003208:Concanavalin A; D011107:Polylysine; D013755:Tetradecanoylphorbol Acetate