Role of MAP kinases p42erk-2/p44erk-1 in cementum-derived attachment-protein-mediated cell attachment.
Cementum-derived attachment protein (CAP) is a collagenous protein which promotes the attachment and spreading of periodontal cell types. We examined the role of the MEK/MAPK pathway in CAP-mediated fibroblast attachment. Human gingival fibroblasts were labeled with 35S-methionine, and the effect of MAP kinase pathway inhibitor PD98059 on attachment and spreading on CAP-coated dishes was examined. Effect on cell proliferation on CAP-coated plates was determined by [3H]-thymidine uptake. Attachment of human gingival fibroblasts to CAP-containing surfaces activated extracellular-signal-regulated kinases (ERK) ERK-2 and ERK-1. In the absence of serum, the ERKs were activated 15 min after attachment, reaching peak levels after 3 hours, and the activity was sustained for at least 12 hours. The enzyme levels were inhibited in cells treated with PD98059. The PD98059 did not significantly affect the kinetics of fibroblast attachment or the number of cells attaching to CAP-coated plates. However, cell spreading was retarded. DNA synthesis as indicated by [3H]-thymidine uptake was not significantly affected. In contrast to PD98059, attachment, spreading, and [3H]-thymidine uptake were inhibited by the protein tyrosine kinase inhibitor genestein. Our results indicate that the MEK/MAPK pathway participates in CAP-mediated fibroblast spreading, but cell attachment and proliferation do not appear to require ERK-2.
Komaki
M
M
Department of Pathology, University of Washington, Seattle 98195, USA.
Kang
M
M
Narayanan
A S
AS
D000818:Animals; D015153:Blotting, Western; D002417:Cattle; D002448:Cell Adhesion / Q000502:physiology*; D015815:Cell Adhesion Molecules / Q000378:metabolism*; D002455:Cell Division; D002465:Cell Movement; D002478:Cells, Cultured; D004247:DNA / Q000096:biosynthesis; D003739:Dental Cementum / Q000201:enzymology*; D004789:Enzyme Activation; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005347:Fibroblasts / Q000201:enzymology; D005419:Flavonoids / Q000494:pharmacology; D005881:Gingiva / Q000166:cytology / Q000201:enzymology*; D006801:Humans; D007334:Insulin-Like Growth Factor I / Q000378:metabolism*; D019950:Mitogen-Activated Protein Kinase 1 / Q000037:antagonists & inhibitors / Q000378:metabolism; D048052:Mitogen-Activated Protein Kinase 3; D020928:Mitogen-Activated Protein Kinases / Q000037:antagonists & inhibitors / Q000378:metabolism*; D015398:Signal Transduction
D015815:Cell Adhesion Molecules; D004791:Enzyme Inhibitors; D005419:Flavonoids; C114460:cementum-derived growth factor, 14-kDa; D007334:Insulin-Like Growth Factor I; D004247:DNA; D019950:Mitogen-Activated Protein Kinase 1; D048052:Mitogen-Activated Protein Kinase 3; D020928:Mitogen-Activated Protein Kinases; C093973:2-(2-amino-3-methoxyphenyl)-4H-1-benzopyran-4-one