Prostaglandin F-induced mitogenesis in MC3T3- E1 osteoblasts: Role of protein kinase-C-mediated tyrosine phosphorylation

Leigh Quarles, Dale M. Haupt, Giora Davidai, John P. Middleton

Research output: Contribution to journalArticle

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Abstract

Prostaglandin F (PGF) stimulates DNA synthesis in osteoblasts through phospholipase-C-dependent increases in intracellular calcium and protein kinase-C (PKC) activity. We present evidence that stimulation of protein tyrosine phosphorylation by PKC is an additional component of the signaling pathways involved in PGF stimulated DNA synthesis in MC3T3- E1 osteoblast-like cells. Mitogenic doses of PGF (42 nM) rapidly induced tyrosine phosphorylation of multiple substrates in these osteoblast-like cells. PGF stimulated tyrosine phosphorylation of new proteins with apparent mol wt of 87, 80, 50, 47, 36, and 33 kilodaltons and up-regulated phosphorylation of preexisting tyrosine components with mol wt of 123, 112, 68, and 56 kilodaltons. Stimulation of PKC by 1.6 μM phorbol 12-myristate 13-acetate mimicked the pattern of PGF -induced protein tyrosine phosphorylation, whereas PKC-deficient cells (induced by overnight pretreatment with 16 μM phorbol 12-myristate 13-acetate) were refractory to PGF -stimulated protein tyrosine phosphorylation and DNA synthesis. The tyrosine kinase inhibitors tyrphostin and genistein blocked PGF -stimulated DNA synthesis and protein tyrosine phosphorylation, and the tyrosine phosphatase inhibitor orthovanadate prolonged PGF -stimulated tyrosine phosphorylation; these findings are consistent with activation of a putative tyrosine kinase. Calcium/calmodulin antagonists also inhibited PGF -stimulated DNA synthesis, but the calciumsignaling pathway played no role in PGF -induced tyrosine phosphorylation. Our findings suggest that cross-talk between receptor-mediated activation of PKC and protein tyrosine phosphorylation is an important distal signaling pathway necessary for PGF -induced DNA synthesis in osteoblast-like cells.

Original languageEnglish (US)
Pages (from-to)1505-1513
Number of pages9
JournalEndocrinology
Volume132
Issue number4
DOIs
StatePublished - Jan 1 1993

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Dinoprost
Osteoblasts
Protein Kinase C
Tyrosine
Phosphorylation
DNA
Protein-Tyrosine Kinases
Proteins
Acetates
Receptor Cross-Talk
Tyrphostins
Calcium
Vanadates
Genistein
Type C Phospholipases
Calmodulin
Phosphoric Monoester Hydrolases

All Science Journal Classification (ASJC) codes

  • Endocrinology

Cite this

Prostaglandin F-induced mitogenesis in MC3T3- E1 osteoblasts : Role of protein kinase-C-mediated tyrosine phosphorylation. / Quarles, Leigh; Haupt, Dale M.; Davidai, Giora; Middleton, John P.

In: Endocrinology, Vol. 132, No. 4, 01.01.1993, p. 1505-1513.

Research output: Contribution to journalArticle

Quarles, Leigh ; Haupt, Dale M. ; Davidai, Giora ; Middleton, John P. / Prostaglandin F-induced mitogenesis in MC3T3- E1 osteoblasts : Role of protein kinase-C-mediated tyrosine phosphorylation. In: Endocrinology. 1993 ; Vol. 132, No. 4. pp. 1505-1513.
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abstract = "Prostaglandin F2α (PGF2α) stimulates DNA synthesis in osteoblasts through phospholipase-C-dependent increases in intracellular calcium and protein kinase-C (PKC) activity. We present evidence that stimulation of protein tyrosine phosphorylation by PKC is an additional component of the signaling pathways involved in PGF2α stimulated DNA synthesis in MC3T3- E1 osteoblast-like cells. Mitogenic doses of PGF2α (42 nM) rapidly induced tyrosine phosphorylation of multiple substrates in these osteoblast-like cells. PGF2α stimulated tyrosine phosphorylation of new proteins with apparent mol wt of 87, 80, 50, 47, 36, and 33 kilodaltons and up-regulated phosphorylation of preexisting tyrosine components with mol wt of 123, 112, 68, and 56 kilodaltons. Stimulation of PKC by 1.6 μM phorbol 12-myristate 13-acetate mimicked the pattern of PGF2α -induced protein tyrosine phosphorylation, whereas PKC-deficient cells (induced by overnight pretreatment with 16 μM phorbol 12-myristate 13-acetate) were refractory to PGF2α -stimulated protein tyrosine phosphorylation and DNA synthesis. The tyrosine kinase inhibitors tyrphostin and genistein blocked PGF2α -stimulated DNA synthesis and protein tyrosine phosphorylation, and the tyrosine phosphatase inhibitor orthovanadate prolonged PGF2α -stimulated tyrosine phosphorylation; these findings are consistent with activation of a putative tyrosine kinase. Calcium/calmodulin antagonists also inhibited PGF2α -stimulated DNA synthesis, but the calciumsignaling pathway played no role in PGF2α -induced tyrosine phosphorylation. Our findings suggest that cross-talk between receptor-mediated activation of PKC and protein tyrosine phosphorylation is an important distal signaling pathway necessary for PGF2α -induced DNA synthesis in osteoblast-like cells.",
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