Skip to main content
NCBI home page
Biochemical Journal logoLink to Biochemical Journal
. 1992 Jul 1;285(Pt 1):229–233. doi: 10.1042/bj2850229

Differential effects of platelet-derived growth factor, serum and bombesin on phospholipase D-mediated hydrolysis of phosphatidylethanolamine in NIH 3T3 fibroblasts.

Z Kiss 1
PMCID: PMC1132770  PMID: 1637304

Abstract

In previous studies, activators of protein kinase C, sphingosine, ATP and various oncogenes were each found to enhance phospholipase D-mediated hydrolysis of phosphatidylethanolamine (PtdEtn) in NIH 3T3 fibroblasts. Here I examined possible stimulation of PtdEtn hydrolysis by various growth-stimulatory agents, including serum, bombesin, platelet-derived growth factor (PDGF), fibroblast growth factor (FGF) and insulin. Treatment of NIH 3T3 fibroblasts, prelabelled with [14C]Etn or [32P]PtdEtn, with PDGF-BB resulted in enhanced formation of [14C]Etn or [32P]phosphatidic acid from the respective labelled cellular pools of PtdEtn. A maximal effect (approximately 3-fold stimulation) on PtdEtn hydrolysis was obtained with 50 ng of PDGF/ml after 5 min of treatment. Phosphatidylcholine (PtdCho) was also hydrolysed, although less extensively than PtdEtn, in PDGF-stimulated cells. PDGF-stimulate hydrolysis of both PtdEtn and PtdCho was prevented by prolonged (30 h) treatment of cells with 400 nM-phorbol 12-myristate 13-acetate (PMA). Similar to PDGF, fetal calf serum (1-10%) also stimulated PtdEtn hydrolysis. However, in contrast to PDGF, the effect of serum on PtdEtn hydrolysis (i) was not diminished by pretreatment with PMA, and (ii) was synergistic with that of PMA after a 1 h incubation. Compared with PDGF and serum, bombesin had less effect on PtdEtn hydrolysis, while FGF and insulin had no effects at all. In contrast to PDGF or serum, bombesin inhibited the effect of PMA on PtdEtn hydrolysis.

Full text

PDF
229

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Besterman J. M., Duronio V., Cuatrecasas P. Rapid formation of diacylglycerol from phosphatidylcholine: a pathway for generation of a second messenger. Proc Natl Acad Sci U S A. 1986 Sep;83(18):6785–6789. doi: 10.1073/pnas.83.18.6785. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Besterman J. M., Watson S. P., Cuatrecasas P. Lack of association of epidermal growth factor-, insulin-, and serum-induced mitogenesis with stimulation of phosphoinositide degradation in BALB/c 3T3 fibroblasts. J Biol Chem. 1986 Jan 15;261(2):723–727. [PubMed] [Google Scholar]
  3. Bierman A. J., Koenderman L., Tool A. J., De Laat S. W. Epidermal growth factor and bombesin differ strikingly in the induction of early responses in Swiss 3T3 cells. J Cell Physiol. 1990 Mar;142(3):441–448. doi: 10.1002/jcp.1041420302. [DOI] [PubMed] [Google Scholar]
  4. Cook S. J., Wakelam M. J. Analysis of the water-soluble products of phosphatidylcholine breakdown by ion-exchange chromatography. Bombesin and TPA (12-O-tetradecanoylphorbol 13-acetate) stimulate choline generation in Swiss 3T3 cells by a common mechanism. Biochem J. 1989 Oct 15;263(2):581–587. doi: 10.1042/bj2630581. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Cook S. J., Wakelam M. J. Hydrolysis of phosphatidylcholine by phospholipase D is a common response to mitogens which stimulate inositol lipid hydrolysis in Swiss 3T3 fibroblasts. Biochim Biophys Acta. 1991 Apr 17;1092(2):265–272. doi: 10.1016/0167-4889(91)90166-u. [DOI] [PubMed] [Google Scholar]
  6. Cook S. J., Wakelam M. J. Hydrolysis of phosphatidylcholine by phospholipase D is a common response to mitogens which stimulate inositol lipid hydrolysis in Swiss 3T3 fibroblasts. Biochim Biophys Acta. 1991 Apr 17;1092(2):265–272. doi: 10.1016/0167-4889(91)90166-u. [DOI] [PubMed] [Google Scholar]
  7. Eichberg J., Hauser G. Stimulation by local anesthetics of the metabolism of acidic phospholipids in the rat pineal gland. Biochem Biophys Res Commun. 1974 Oct 23;60(4):1460–1467. doi: 10.1016/0006-291x(74)90362-3. [DOI] [PubMed] [Google Scholar]
  8. Fisher G. J., Henderson P. A., Voorhees J. J., Baldassare J. J. Epidermal growth factor-induced hydrolysis of phosphatidylcholine by phospholipase D and phospholipase C in human dermal fibroblasts. J Cell Physiol. 1991 Feb;146(2):309–317. doi: 10.1002/jcp.1041460216. [DOI] [PubMed] [Google Scholar]
  9. Heslop J. P., Blakeley D. M., Brown K. D., Irvine R. F., Berridge M. J. Effects of bombesin and insulin on inositol (1,4,5)trisphosphate and inositol (1,3,4)trisphosphate formation in Swiss 3T3 cells. Cell. 1986 Dec 5;47(5):703–709. doi: 10.1016/0092-8674(86)90513-1. [DOI] [PubMed] [Google Scholar]
  10. Hii C. S., Edwards Y. S., Murray A. W. Phorbol ester-stimulated hydrolysis of phosphatidylcholine and phosphatidylethanolamine by phospholipase D in HeLa cells. Evidence that the basal turnover of phosphoglycerides does not involve phospholipase D. J Biol Chem. 1991 Oct 25;266(30):20238–20243. [PubMed] [Google Scholar]
  11. Hovis J. G., Stumpo D. J., Halsey D. L., Blackshear P. J. Effects of mitogens on ornithine decarboxylase activity and messenger RNA levels in normal and protein kinase C-deficient NIH-3T3 fibroblasts. J Biol Chem. 1986 Aug 5;261(22):10380–10386. [PubMed] [Google Scholar]
  12. Kiss Z., Anderson W. B. ATP stimulates the hydrolysis of phosphatidylethanolamine in NIH 3T3 cells. Potentiating effects of guanosine triphosphates and sphingosine. J Biol Chem. 1990 May 5;265(13):7345–7350. [PubMed] [Google Scholar]
  13. Kiss Z., Anderson W. B. Alcohols selectively stimulate phospholipase D-mediated hydrolysis of phosphatidylethanolamine in NIH 3T3 cells. FEBS Lett. 1989 Oct 23;257(1):45–48. doi: 10.1016/0014-5793(89)81782-x. [DOI] [PubMed] [Google Scholar]
  14. Kiss Z., Anderson W. B. Phorbol ester stimulates the hydrolysis of phosphatidylethanolamine in leukemic HL-60, NIH 3T3, and baby hamster kidney cells. J Biol Chem. 1989 Jan 25;264(3):1483–1487. [PubMed] [Google Scholar]
  15. Kiss Z., Chattopadhyay J., Pettit G. R. Stimulation of phosphatidylcholine synthesis by activators of protein kinase C is dissociable from increased phospholipid hydrolysis. Biochem J. 1991 Jan 1;273(Pt 1):189–194. doi: 10.1042/bj2730189. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Kiss Z., Crilly K. S. Adenosine triphosphate (ATP) and other nucleotides stimulate the hydrolysis of phosphatidylethanolamine in intact fibroblasts. Lipids. 1991 Oct;26(10):777–780. doi: 10.1007/BF02536157. [DOI] [PubMed] [Google Scholar]
  17. Kiss Z. Determination of phospholipase D-mediated hydrolysis of phosphatidylethanolamine. Lipids. 1991 Apr;26(4):321–326. doi: 10.1007/BF02537144. [DOI] [PubMed] [Google Scholar]
  18. Kiss Z., Rapp U. R., Pettit G. R., Anderson W. B. Phorbol ester and bryostatin differentially regulate the hydrolysis of phosphatidylethanolamine in Ha-ras- and raf-oncogene-transformed NIH 3T3 cells. Biochem J. 1991 Jun 1;276(Pt 2):505–509. doi: 10.1042/bj2760505. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Koul O., Hauser G. Modulation of rat brain cytosolic phosphatidate phosphohydrolase: effect of cationic amphiphilic drugs and divalent cations. Arch Biochem Biophys. 1987 Mar;253(2):453–461. doi: 10.1016/0003-9861(87)90199-8. [DOI] [PubMed] [Google Scholar]
  20. L'Allemain G., Pouysségur EGF and insulin action in fibroblasts. Evidence that phosphoinositide hydrolysis is not an essential mitogenic signalling pathway. FEBS Lett. 1986 Mar 3;197(1-2):344–348. doi: 10.1016/0014-5793(86)80354-4. [DOI] [PubMed] [Google Scholar]
  21. Larrodera P., Cornet M. E., Diaz-Meco M. T., Lopez-Barahona M., Diaz-Laviada I., Guddal P. H., Johansen T., Moscat J. Phospholipase C-mediated hydrolysis of phosphatidylcholine is an important step in PDGF-stimulated DNA synthesis. Cell. 1990 Jun 15;61(6):1113–1120. doi: 10.1016/0092-8674(90)90074-o. [DOI] [PubMed] [Google Scholar]
  22. Lavie Y., Liscovitch M. Activation of phospholipase D by sphingoid bases in NG108-15 neural-derived cells. J Biol Chem. 1990 Mar 5;265(7):3868–3872. [PubMed] [Google Scholar]
  23. Lopez-Rivas A., Mendoza S. A., Nånberg E., Sinnett-Smith J., Rozengurt E. Ca2+-mobilizing actions of platelet-derived growth factor differ from those of bombesin and vasopressin in Swiss 3T3 mouse cells. Proc Natl Acad Sci U S A. 1987 Aug;84(16):5768–5772. doi: 10.1073/pnas.84.16.5768. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Magnaldo I., L'Allemain G., Chambard J. C., Moenner M., Barritault D., Pouysségur J. The mitogenic signaling pathway of fibroblast growth factor is not mediated through polyphosphoinositide hydrolysis and protein kinase C activation in hamster fibroblasts. J Biol Chem. 1986 Dec 25;261(36):16916–16922. [PubMed] [Google Scholar]
  25. Pappu A. S., Hauser G. Propranolol-induced inhibition of rat brain cytoplasmic phosphatidate phosphohydrolase. Neurochem Res. 1983 Dec;8(12):1565–1575. doi: 10.1007/BF00964158. [DOI] [PubMed] [Google Scholar]
  26. Pessin M. S., Baldassare J. J., Raben D. M. Molecular species analysis of mitogen-stimulated 1,2-diglycerides in fibroblasts. Comparison of alpha-thrombin, epidermal growth factor, and platelet-derived growth factor. J Biol Chem. 1990 May 15;265(14):7959–7966. [PubMed] [Google Scholar]
  27. Price B. D., Morris J. D., Hall A. Stimulation of phosphatidylcholine breakdown and diacylglycerol production by growth factors in Swiss-3T3 cells. Biochem J. 1989 Dec 1;264(2):509–515. doi: 10.1042/bj2640509. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Raben D. M., Pessin M. S., Rangan L. A., Wright T. M. Kinetic and molecular species analyses of mitogen-induced increases in diglycerides: evidence for stimulated hydrolysis of phosphoinositides and phosphatidylcholine. J Cell Biochem. 1990 Oct;44(2):117–125. doi: 10.1002/jcb.240440206. [DOI] [PubMed] [Google Scholar]
  29. Rodriguez-Pena A., Rozengurt E. Disappearance of Ca2+-sensitive, phospholipid-dependent protein kinase activity in phorbol ester-treated 3T3 cells. Biochem Biophys Res Commun. 1984 May 16;120(3):1053–1059. doi: 10.1016/s0006-291x(84)80213-2. [DOI] [PubMed] [Google Scholar]
  30. Vicentini L. M., Villereal M. L. Serum, bradykinin and vasopressin stimulate release of inositol phosphates from human fibroblasts. Biochem Biophys Res Commun. 1984 Sep 17;123(2):663–670. doi: 10.1016/0006-291x(84)90280-8. [DOI] [PubMed] [Google Scholar]
  31. Wang P., Anthes J. C., Siegel M. I., Egan R. W., Billah M. M. Existence of cytosolic phospholipase D. Identification and comparison with membrane-bound enzyme. J Biol Chem. 1991 Aug 15;266(23):14877–14880. [PubMed] [Google Scholar]
  32. Wright T. M., Rangan L. A., Shin H. S., Raben D. M. Kinetic analysis of 1,2-diacylglycerol mass levels in cultured fibroblasts. Comparison of stimulation by alpha-thrombin and epidermal growth factor. J Biol Chem. 1988 Jul 5;263(19):9374–9380. [PubMed] [Google Scholar]
  33. Wright T. M., Shin H. S., Raben D. M. Sustained increase in 1,2-diacylglycerol precedes DNA synthesis in epidermal-growth-factor-stimulated fibroblasts. Evidence for stimulated phosphatidylcholine hydrolysis. Biochem J. 1990 Apr 15;267(2):501–507. doi: 10.1042/bj2670501. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Biochemical Journal are provided here courtesy of The Biochemical Society

RESOURCES