Transforming growth factor β enhances parathyroid hormone stimulation of adenylate cyclase in clonal osteoblast-like cells

Gloria E. Gutierrez, Gregory R. Mundy, David R. Manning, Erik L. Hewlett, Michael S. Katz

Research output: Contribution to journalArticle

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Abstract

The effects of transforming growth factor β (TGFβ) on parathyroid hormone (PTH)-responsive adenylate cyclase were examined in clonal rat osteosarcoma cells (UMR-106) with the osteoblast phenotype. Purified TGFβ incubated with UMR-106 cells for 48 hr produced a concentration-dependent increase in PTH stimulation of adenylate cyclase, with maximal increase in PTH response (37%) occurring at 1 ng/ml TGFβ. TGFβ also enhanced receptor-mediated activation of adenylate cyclase by isoproterenol and prostaglandin E2 (PGE2) and nonreceptor-mediated enzyme activation by cholera toxin and forskolin. In cells in which PTH-stimulated adenylate cyclase activity was augmented by treatment with pertussis toxin, the incremental increase in PTH response produced by TGFβ was reduced by 33%. However, TGFβ neither mimicked nor altered the ability of pertussis toxin to catalyze the ADP-ribosylation of a 41,000-Da protein, presumably the α subunit of the inhibitory guanine nucleotide-binding regulatory component (Gi) of adenylate cyclase, in cholate-extracted UMR-106 cell membranes. TGFβ also had no effect on the levels of α or β subunits of Gi, as assessed by immunotransfer blotting. In time course studies, brief (≤30 min) exposure of cells to TGFβ during early culture was sufficient to increase PTH response but only after exposed cells were subsequently allowed to grow for prolonged periods. TGFβ enhancement of PTH and isoproterenol responses was blocked by prior treatment of cells with cycloheximide but not indomethacin. The results suggest that TGFβ enhances PTH response in osteoblast-like cells by action(s) exerted at nonreceptor components of adenylate cyclase. The effect of TGFβ may involve Gi, although in a manner unrelated to either pertussis toxin-catalyzed ADP-ribosylation of the α subunit of Gi or changes in levels of Gi subunits. The regulatory action of TGFβ on adenylate cyclase is likely to be mediated by the rapid generation of cellular signals excluding prostaglandins, followed by a prolonged sequence of events involving protein synthesis. These observations suggest a mechanism by which TGFβ may regulate osteoblast responses to systemic hormones.

Original languageEnglish (US)
Pages (from-to)438-447
Number of pages10
JournalJournal of Cellular Physiology
Volume144
Issue number3
StatePublished - Sep 1990

Fingerprint

Osteoblasts
Transforming Growth Factors
Parathyroid Hormone
Adenylyl Cyclases
Pertussis Toxin
Isoproterenol
Adenosine Diphosphate
Chemical activation
Cholates
Time and motion study
Enzyme Activation
Guanine Nucleotides
Cholera Toxin
Protein Subunits
Colforsin
Osteosarcoma
Cell membranes
Cycloheximide
Dinoprostone
Indomethacin

ASJC Scopus subject areas

  • Clinical Biochemistry
  • Cell Biology
  • Physiology

Cite this

Gutierrez, G. E., Mundy, G. R., Manning, D. R., Hewlett, E. L., & Katz, M. S. (1990). Transforming growth factor β enhances parathyroid hormone stimulation of adenylate cyclase in clonal osteoblast-like cells. Journal of Cellular Physiology, 144(3), 438-447.

Transforming growth factor β enhances parathyroid hormone stimulation of adenylate cyclase in clonal osteoblast-like cells. / Gutierrez, Gloria E.; Mundy, Gregory R.; Manning, David R.; Hewlett, Erik L.; Katz, Michael S.

In: Journal of Cellular Physiology, Vol. 144, No. 3, 09.1990, p. 438-447.

Research output: Contribution to journalArticle

Gutierrez, GE, Mundy, GR, Manning, DR, Hewlett, EL & Katz, MS 1990, 'Transforming growth factor β enhances parathyroid hormone stimulation of adenylate cyclase in clonal osteoblast-like cells', Journal of Cellular Physiology, vol. 144, no. 3, pp. 438-447.
Gutierrez, Gloria E. ; Mundy, Gregory R. ; Manning, David R. ; Hewlett, Erik L. ; Katz, Michael S. / Transforming growth factor β enhances parathyroid hormone stimulation of adenylate cyclase in clonal osteoblast-like cells. In: Journal of Cellular Physiology. 1990 ; Vol. 144, No. 3. pp. 438-447.
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abstract = "The effects of transforming growth factor β (TGFβ) on parathyroid hormone (PTH)-responsive adenylate cyclase were examined in clonal rat osteosarcoma cells (UMR-106) with the osteoblast phenotype. Purified TGFβ incubated with UMR-106 cells for 48 hr produced a concentration-dependent increase in PTH stimulation of adenylate cyclase, with maximal increase in PTH response (37{\%}) occurring at 1 ng/ml TGFβ. TGFβ also enhanced receptor-mediated activation of adenylate cyclase by isoproterenol and prostaglandin E2 (PGE2) and nonreceptor-mediated enzyme activation by cholera toxin and forskolin. In cells in which PTH-stimulated adenylate cyclase activity was augmented by treatment with pertussis toxin, the incremental increase in PTH response produced by TGFβ was reduced by 33{\%}. However, TGFβ neither mimicked nor altered the ability of pertussis toxin to catalyze the ADP-ribosylation of a 41,000-Da protein, presumably the α subunit of the inhibitory guanine nucleotide-binding regulatory component (Gi) of adenylate cyclase, in cholate-extracted UMR-106 cell membranes. TGFβ also had no effect on the levels of α or β subunits of Gi, as assessed by immunotransfer blotting. In time course studies, brief (≤30 min) exposure of cells to TGFβ during early culture was sufficient to increase PTH response but only after exposed cells were subsequently allowed to grow for prolonged periods. TGFβ enhancement of PTH and isoproterenol responses was blocked by prior treatment of cells with cycloheximide but not indomethacin. The results suggest that TGFβ enhances PTH response in osteoblast-like cells by action(s) exerted at nonreceptor components of adenylate cyclase. The effect of TGFβ may involve Gi, although in a manner unrelated to either pertussis toxin-catalyzed ADP-ribosylation of the α subunit of Gi or changes in levels of Gi subunits. The regulatory action of TGFβ on adenylate cyclase is likely to be mediated by the rapid generation of cellular signals excluding prostaglandins, followed by a prolonged sequence of events involving protein synthesis. These observations suggest a mechanism by which TGFβ may regulate osteoblast responses to systemic hormones.",
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