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.
Abrieu A., Fisher D., Simon M.-N, Doree M., Picard A. (1997). MAPK inactivation is required for the G2 to M-phase transition of the first mitotic cell cycle. EMBO J16:6407-6413.
2.
Alessi DR, Cuenda A., Cohen P., Dudley DT, Saltiel AR (1995). PD 98059 is a specific inhibitor of the activation of mitogen-activated protein kinase kinase in vitro and in vivo. J Biol Chem270 :27489-27494.
3.
Assoian RK, Zhu X. (1997). Cell anchorage and the cytoskeleton as partners in growth factor dependent cell cycle progression. Curr Opin Cell Biol9:93-98.
4.
Arzate H., Olson SW, Page RC, Gown AM, Narayanan AS (1992). Production of a monoclonal antibody to an attachment protein derived from human cementum. FASEB J6:2990-2995.
5.
Bar-Kana I., Savion N., Narayanan AS, Pitaru S. (1998). Cementum attachment protein manifestation is restricted to the mineralized tissue forming cells of the periodontium. Eur J Oral Sci106(Suppl 1):357-364.
6.
Bartold PM, McCulloch Cag, Narayanan AS, Pitaru S. (2000). Tissue engineering. A new paradigm for periodontal regeneration based on molecular and cell biology. Periodontology200024:253-269.
7.
Bennett AM, Tonks NK (1997). Regulation of distinct stages of skeletal muscle differentiation by mitogen-activated protein kinases. Science278:1288-1291.
8.
Bohmer R.-M, Scharf E., Assoian RK (1996). Cytoskeletal integrity is required throughout the mitogen stimulation phase of the cell cycle and mediates the anchorage-dependent expression of cyclin D1. Mol Biol Cell7: 101-111.
9.
Clark Raf (1996). The molecular and cellular biology of wound repair . 2nd ed. New York: Plenum Press.
10.
Dudley DT, Pang L., Decker SJ, Bridges AJ, Saltiel AR (1995). A synthetic inhibitor of the mitogen-activated protein kinase cascade. Proc Natl Acad Sci USA92:7686-7689.
11.
Fukunaga K., Miyamoto E. (1998). Role of MAP kinase in neurons. Mol Neurobiol16:79-95.
12.
Gumbiner BM (1996). Cell adhesion: the molecular basis of tissue architecture and morphogenesis. Cell84:345-357.
13.
Ivanovski S., Komaki M.Bartold PM, Narayanan AS (1999). Periodontal-derived cells attach to cementum attachment protein via alpha5betal integrin. J Periodont Res34:154-159.
14.
Juliano RL, Haskill S. (1993). Signal transduction from the extracellular matrix . J Cell Biol120:577-585.
15.
Karin M. (1995). The regulation of AP-1 activity by mitogen activated protein kinases. J Biol Chem270:16483-16486.
16.
Lavoie JN, L'Allemain G., Brunet A., Muller R., Pouyssegur J. (1996). Cyclin D1 expression is regulated positively by the p42/p44MAPK and negatively by the p38/HOGMAPK pathway. J Biol Chem271:20608-20616.
17.
Lewis TS, Shapiro PS, Ahn NG (1998). Signal transduction through MAP kinase cascades . Adv Cancer Res74:49-139.
18.
Liu HW, Yacobi R., Savion N., Narayanan AS, Pitaru S. (1997). A collagenous cementum-derived attachment protein is a marker for progenitors of the mineralized tissue-forming cell lineage of the periodontal ligament. J Bone Miner Res12:1691-1699.
19.
Mansour SJ, Matten WT, Hermann AS, Candia JM, Rong S., Fukasawa K., et al. (1994). Transformation of mammalian cells by constitutively active MAP kinase. Science265:966-970.
20.
Miyamoto S., Akiyama SK, Yamada KM (1995). Synergistic roles for receptor occupancy and aggregation in integrin transmembrane function. Science267:883-885.
21.
Morino N., Mimura T., Hamasaki K., Tobe K., Ueki K., Kikuchi K., et al. (1995). Matrix/integrin interaction activates the mitogen-activated protein kinase, p44erk-1 and p42erk-2. J Biol Chem270:269-273.
22.
Olson S., Arzate H., Narayanan AS, Page RC ( 1991). Cell attachment activity of cementum proteins and mechanism of endotoxin inhibition. J Dent Res70:1272-1277.
23.
Pitaru S., McCulloch CA, Narayanan SA (1994). Cellular origins and differentiation control mechanisms during periodontal development and wound healing. J Periodont Res29:81-94.
24.
Pitaru S., Narayanan SA, Olson S., Savion N., Hekmati H., Alt I., et al. (1995). Specific cementum attachment protein enhances selectively the attachment and migration of periodontal cells to root surfaces . J Periodont Res30:360-368.
25.
Racke FK, Lewandowska K., Goueli S., Goldfarb AN (1997). Sustained activation of the extracellular signal-regulated kinase/mitogenactivated protein kinase pathway is required for megakaryocytic differentiation of K562 Cells. J Biol Chem272:23366-23370.
26.
Ramakrishnan M., Musa NL, Li J., Liu PT, Pestell RG, Hershenson MB (1998). Catalytic activation of extracellular signal-regulated kinases induces cyclin D1 expression in primary tracheal myocytes. Am J Resp Cell Mol Biol18:736-740.
27.
Saito M., Narayanan AS (1999). Signaling reactions induced in human fibroblasts during adhesion to cementum-derived attachment protein. J Bone Miner Res14:65-72.
28.
Sastry SK, Horwitz AF (1996). Adhesion-growth factor interactions during differentiation: an integrated biological response. Dev Biol180:455-467.
29.
Seger R., Krebs EG (1995). The MAPK signaling cascade. FASEB J9:726-735.
30.
Weber JD, Hu W., Jefcoat SC Jr, Raben DM, Baldassare JJ (1997). Ras-stimulated extracellular signal-related kinase 1 and RhoA activities coordinate platelet-derived growth factor-induced G1 progression through the independent regulation of cyclin D1 and p27. J Biol Chem272:32966-32971.
31.
Wu D., Ikezawa K., Parker T., Saito M., Narayanan AS (1996). Characterization of a collagenous cementum-derived attachment protein. J Bone Miner Res11:686-692.
32.
Yamada KM, Miyamoto S. (1995). Integrin transmembrane signaling and cytoskeletal control. Curr Opin Cell Biol7:681-689.
33.
Yonemura K., Raines EW, Ahn NG, Narayanan AS (1993). Mitogenic signaling mechanisms of human cementum-derived growth factors. J Biol Chem268:26120-26126.
34.
Zhu X., Assoian RK (1995). Integrin dependent activation of MAP kinase: a link to shape-dependent cell proliferation. Mol Biol Cell6:273-282.
