The human 5-HT(1A) receptor expressed in HeLa cells stimulates sodium-dependent phosphate uptake via protein kianse C

J. R. Raymond, A. Fargin, J. P. Middleton, J. M. Graff, D. McNeill Haupt, M. G. Caron, R. J. Lefkowitz, V. W. Dennis

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Abstract

Regulation of phosphate uptake was studied in HeLa cell lines after transfection with DNA encoding the human 5-HT(1A) receptor. Phosphate uptake was saturable and >90% sodium-depEndent, with V(max) ~ 4.3 nmol/mg protein/3 min and K(m) ~ 0.4 mM phosphate. Activators of protein kinase A decreased V(max) by ~ 30-35% without changing K(m). Treatment with 5-HT or the 5-HT(1A)-specific agonist 8-OH-2-(di-n-propylamino)1,2,3,4-tetrahydronaphthalene increased V(max) ~ 40% without affecting K(m). This effect was blocked by pretreatment with the 5-HT1 antagonists, methiothepine and spiperone, or pertussis toxin. Surprisingly, the stimulation was not secondary to an inhibition of adenylyl cyclase because 5-HT stimulated phosphate uptake ~ 20% in the presence of 1 mM 8-Br-cAMP. Rather, the primary pathway linked to the stimulation of phosphate uptake involved activation of protein kinase C because (i) 5-HT measurably activated protein kinase C in these cells, (ii) activators of protein kinase C (phorbol esters and diacylglycerol analogues) stimulated phosphate uptake in these cells (iii) the half-maximal doses for 5-HT-induced phosphatidylinositol hydrolysis and stimulation of phosphate uptake were virtually equivalent, and both effects were equally sensitive to pertussis toxin, and (iv) the stimulation was markedly attenuated in cells made deficient in protein kinase C. These results demonstrate that the stimulation of phosphatidylinositol hydrolysis by the 5-HT(1A) receptor can generate physiologically measurable effects on cellular transport and suggest that such accessory pathways may play a prominent role in signal transduction.

Original languageEnglish (US)
Pages (from-to)21943-21950
Number of pages8
JournalJournal of Biological Chemistry
Volume264
Issue number36
StatePublished - 1989

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Receptor, Serotonin, 5-HT1A
Protein C
HeLa Cells
Sodium
Phosphates
Serotonin
Protein Kinase C
Pertussis Toxin
Phosphatidylinositols
Hydrolysis
Methiothepin
Serotonin 5-HT1 Receptor Antagonists
Spiperone
Signal transduction
Diglycerides
Accessories
Phorbol Esters
Cyclic AMP-Dependent Protein Kinases
sodium phosphate
Transfection

ASJC Scopus subject areas

  • Biochemistry

Cite this

Raymond, J. R., Fargin, A., Middleton, J. P., Graff, J. M., McNeill Haupt, D., Caron, M. G., ... Dennis, V. W. (1989). The human 5-HT(1A) receptor expressed in HeLa cells stimulates sodium-dependent phosphate uptake via protein kianse C. Journal of Biological Chemistry, 264(36), 21943-21950.

The human 5-HT(1A) receptor expressed in HeLa cells stimulates sodium-dependent phosphate uptake via protein kianse C. / Raymond, J. R.; Fargin, A.; Middleton, J. P.; Graff, J. M.; McNeill Haupt, D.; Caron, M. G.; Lefkowitz, R. J.; Dennis, V. W.

In: Journal of Biological Chemistry, Vol. 264, No. 36, 1989, p. 21943-21950.

Research output: Contribution to journalArticle

Raymond, JR, Fargin, A, Middleton, JP, Graff, JM, McNeill Haupt, D, Caron, MG, Lefkowitz, RJ & Dennis, VW 1989, 'The human 5-HT(1A) receptor expressed in HeLa cells stimulates sodium-dependent phosphate uptake via protein kianse C', Journal of Biological Chemistry, vol. 264, no. 36, pp. 21943-21950.
Raymond, J. R. ; Fargin, A. ; Middleton, J. P. ; Graff, J. M. ; McNeill Haupt, D. ; Caron, M. G. ; Lefkowitz, R. J. ; Dennis, V. W. / The human 5-HT(1A) receptor expressed in HeLa cells stimulates sodium-dependent phosphate uptake via protein kianse C. In: Journal of Biological Chemistry. 1989 ; Vol. 264, No. 36. pp. 21943-21950.
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abstract = "Regulation of phosphate uptake was studied in HeLa cell lines after transfection with DNA encoding the human 5-HT(1A) receptor. Phosphate uptake was saturable and >90{\%} sodium-depEndent, with V(max) ~ 4.3 nmol/mg protein/3 min and K(m) ~ 0.4 mM phosphate. Activators of protein kinase A decreased V(max) by ~ 30-35{\%} without changing K(m). Treatment with 5-HT or the 5-HT(1A)-specific agonist 8-OH-2-(di-n-propylamino)1,2,3,4-tetrahydronaphthalene increased V(max) ~ 40{\%} without affecting K(m). This effect was blocked by pretreatment with the 5-HT1 antagonists, methiothepine and spiperone, or pertussis toxin. Surprisingly, the stimulation was not secondary to an inhibition of adenylyl cyclase because 5-HT stimulated phosphate uptake ~ 20{\%} in the presence of 1 mM 8-Br-cAMP. Rather, the primary pathway linked to the stimulation of phosphate uptake involved activation of protein kinase C because (i) 5-HT measurably activated protein kinase C in these cells, (ii) activators of protein kinase C (phorbol esters and diacylglycerol analogues) stimulated phosphate uptake in these cells (iii) the half-maximal doses for 5-HT-induced phosphatidylinositol hydrolysis and stimulation of phosphate uptake were virtually equivalent, and both effects were equally sensitive to pertussis toxin, and (iv) the stimulation was markedly attenuated in cells made deficient in protein kinase C. These results demonstrate that the stimulation of phosphatidylinositol hydrolysis by the 5-HT(1A) receptor can generate physiologically measurable effects on cellular transport and suggest that such accessory pathways may play a prominent role in signal transduction.",
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T1 - The human 5-HT(1A) receptor expressed in HeLa cells stimulates sodium-dependent phosphate uptake via protein kianse C

AU - Raymond, J. R.

AU - Fargin, A.

AU - Middleton, J. P.

AU - Graff, J. M.

AU - McNeill Haupt, D.

AU - Caron, M. G.

AU - Lefkowitz, R. J.

AU - Dennis, V. W.

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N2 - Regulation of phosphate uptake was studied in HeLa cell lines after transfection with DNA encoding the human 5-HT(1A) receptor. Phosphate uptake was saturable and >90% sodium-depEndent, with V(max) ~ 4.3 nmol/mg protein/3 min and K(m) ~ 0.4 mM phosphate. Activators of protein kinase A decreased V(max) by ~ 30-35% without changing K(m). Treatment with 5-HT or the 5-HT(1A)-specific agonist 8-OH-2-(di-n-propylamino)1,2,3,4-tetrahydronaphthalene increased V(max) ~ 40% without affecting K(m). This effect was blocked by pretreatment with the 5-HT1 antagonists, methiothepine and spiperone, or pertussis toxin. Surprisingly, the stimulation was not secondary to an inhibition of adenylyl cyclase because 5-HT stimulated phosphate uptake ~ 20% in the presence of 1 mM 8-Br-cAMP. Rather, the primary pathway linked to the stimulation of phosphate uptake involved activation of protein kinase C because (i) 5-HT measurably activated protein kinase C in these cells, (ii) activators of protein kinase C (phorbol esters and diacylglycerol analogues) stimulated phosphate uptake in these cells (iii) the half-maximal doses for 5-HT-induced phosphatidylinositol hydrolysis and stimulation of phosphate uptake were virtually equivalent, and both effects were equally sensitive to pertussis toxin, and (iv) the stimulation was markedly attenuated in cells made deficient in protein kinase C. These results demonstrate that the stimulation of phosphatidylinositol hydrolysis by the 5-HT(1A) receptor can generate physiologically measurable effects on cellular transport and suggest that such accessory pathways may play a prominent role in signal transduction.

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