Skip to main content
The Journal of Cell Biology logoLink to The Journal of Cell Biology
. 1991 May 2;113(4):943–950. doi: 10.1083/jcb.113.4.943

Mastoparan, a novel mitogen for Swiss 3T3 cells, stimulates pertussis toxin-sensitive arachidonic acid release without inositol phosphate accumulation

PMCID: PMC2288979  PMID: 1709171

Abstract

Mastoparan, a basic tetradecapeptide isolated from wasp venom, is a novel mitogen for Swiss 3T3 cells. This peptide induced DNA synthesis in synergy with insulin in a concentration-dependent manner; half- maximum and maximum responses were achieved at 14 and 17 microM, respectively. Mastoparan also stimulated DNA synthesis in the presence of other growth promoting factors including bombesin, insulin-like growth factor-1, and platelet-derived growth factor. The synergistic mitogenic stimulation by mastoparan can be dissociated from activation of phospholipase C. Mastoparan did not stimulate phosphoinositide breakdown, Ca2+ mobilization or protein kinase C-mediated phosphorylation of a major cellular substrate or transmodulation of the epidermal growth factor receptor. In contrast, mastoparan stimulated arachidonic acid release, prostaglandin E2 production, and enhanced cAMP accumulation in the presence of forskolin. These responses were inhibited by prior treatment with pertussis toxin. Hence, mastoparan stimulates arachidonic acid release via a pertussis toxin-sensitive G protein in Swiss 3T3 cells. Arachidonic acid, like mastoparan, stimulated DNA synthesis in the presence of insulin. The ability of mastoparan to stimulate mitogenesis was reduced by pertussis toxin treatment. These results demonstrate, for the first time, that mastoparan stimulates reinitiation of DNA synthesis in Swiss 3T3 cells and indicate that this peptide may be a useful probe to elucidate signal transduction mechanisms in mitogenesis.

Full Text

The Full Text of this article is available as a PDF (988.4 KB).

Selected References

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

  1. Argiolas A., Pisano J. J. Facilitation of phospholipase A2 activity by mastoparans, a new class of mast cell degranulating peptides from wasp venom. J Biol Chem. 1983 Nov 25;258(22):13697–13702. [PubMed] [Google Scholar]
  2. Aridor M., Traub L. M., Sagi-Eisenberg R. Exocytosis in mast cells by basic secretagogues: evidence for direct activation of GTP-binding proteins. J Cell Biol. 1990 Sep;111(3):909–917. doi: 10.1083/jcb.111.3.909. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Axelrod J. Receptor-mediated activation of phospholipase A2 and arachidonic acid release in signal transduction. Biochem Soc Trans. 1990 Aug;18(4):503–507. doi: 10.1042/bst0180503. [DOI] [PubMed] [Google Scholar]
  4. Berridge M. J., Dawson R. M., Downes C. P., Heslop J. P., Irvine R. F. Changes in the levels of inositol phosphates after agonist-dependent hydrolysis of membrane phosphoinositides. Biochem J. 1983 May 15;212(2):473–482. doi: 10.1042/bj2120473. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Camoratto A. M., Grandison L. Evidence supporting a correlation between arachidonic acid release and prolactin secretion from GH3 cells. Endocrinology. 1985 Apr;116(4):1506–1513. doi: 10.1210/endo-116-4-1506. [DOI] [PubMed] [Google Scholar]
  6. Coffer A., Fabregat I., Sinnett-Smith J., Rozengurt E. Solubilization of the bombesin receptor from Swiss 3T3 cell membranes. Functional association to a guanine nucleotide regulatory protein. FEBS Lett. 1990 Apr 9;263(1):80–84. doi: 10.1016/0014-5793(90)80710-z. [DOI] [PubMed] [Google Scholar]
  7. Dicker P., Rozengurt E. Phorbol esters and vasopressin stimulate DNA synthesis by a common mechanism. Nature. 1980 Oct 16;287(5783):607–612. doi: 10.1038/287607a0. [DOI] [PubMed] [Google Scholar]
  8. Erusalimsky J. D., Friedberg I., Rozengurt E. Bombesin, diacylglycerols, and phorbol esters rapidly stimulate the phosphorylation of an Mr = 80,000 protein kinase C substrate in permeabilized 3T3 cells. Effect of guanine nucleotides. J Biol Chem. 1988 Dec 15;263(35):19188–19194. [PubMed] [Google Scholar]
  9. Erusalimsky J. D., Rozengurt E. Vasopressin rapidly stimulates protein kinase C in digitonin-permeabilized Swiss 3T3 cells: involvement of a pertussis toxin-insensitive guanine nucleotide binding protein. J Cell Physiol. 1989 Nov;141(2):253–261. doi: 10.1002/jcp.1041410204. [DOI] [PubMed] [Google Scholar]
  10. Gilman A. G. G proteins: transducers of receptor-generated signals. Annu Rev Biochem. 1987;56:615–649. doi: 10.1146/annurev.bi.56.070187.003151. [DOI] [PubMed] [Google Scholar]
  11. 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]
  12. Higashijima T., Burnier J., Ross E. M. Regulation of Gi and Go by mastoparan, related amphiphilic peptides, and hydrophobic amines. Mechanism and structural determinants of activity. J Biol Chem. 1990 Aug 25;265(24):14176–14186. [PubMed] [Google Scholar]
  13. Higashijima T., Uzu S., Nakajima T., Ross E. M. Mastoparan, a peptide toxin from wasp venom, mimics receptors by activating GTP-binding regulatory proteins (G proteins). J Biol Chem. 1988 May 15;263(14):6491–6494. [PubMed] [Google Scholar]
  14. Higashijima T., Wakamatsu K., Takemitsu M., Fujino M., Nakajima T., Miyazawa T. Conformational change of mastoparan from wasp venom on binding with phospholipid membrane. FEBS Lett. 1983 Feb 21;152(2):227–230. doi: 10.1016/0014-5793(83)80385-8. [DOI] [PubMed] [Google Scholar]
  15. Hirai Y., Yasuhara T., Yoshida H., Nakajima T., Fujino M., Kitada C. A new mast cell degranulating peptide "mastoparan" in the venom of Vespula lewisii. Chem Pharm Bull (Tokyo) 1979 Aug;27(8):1942–1944. doi: 10.1248/cpb.27.1942. [DOI] [PubMed] [Google Scholar]
  16. Kurihara H., Kitajima K., Senda T., Fujita H., Nakajima T. Multigranular exocytosis induced by phospholipase A2-activators, melittin and mastoparan, in rat anterior pituitary cells. Cell Tissue Res. 1986;243(2):311–316. doi: 10.1007/BF00251045. [DOI] [PubMed] [Google Scholar]
  17. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  18. 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]
  19. Mehmet H., Nånberg E., Lehmann W., Murray M. J., Rozengurt E. Early signals in the mitogenic response of Swiss 3T3 cells: a comparative study of purified PDGF homodimers. Growth Factors. 1990;3(2):83–95. doi: 10.3109/08977199009108271. [DOI] [PubMed] [Google Scholar]
  20. Millar J. B., Rozengurt E. Arachidonic acid release by bombesin. A novel postreceptor target for heterologous mitogenic desensitization. J Biol Chem. 1990 Nov 15;265(32):19973–19979. [PubMed] [Google Scholar]
  21. Millar J. B., Rozengurt E. Bombesin enhancement of cAMP accumulation in Swiss 3T3 cells: evidence of a dual mechanism of action. J Cell Physiol. 1988 Nov;137(2):214–222. doi: 10.1002/jcp.1041370203. [DOI] [PubMed] [Google Scholar]
  22. Mousli M., Bronner C., Bueb J. L., Tschirhart E., Gies J. P., Landry Y. Activation of rat peritoneal mast cells by substance P and mastoparan. J Pharmacol Exp Ther. 1989 Jul;250(1):329–335. [PubMed] [Google Scholar]
  23. Mousli M., Bronner C., Landry Y., Bockaert J., Rouot B. Direct activation of GTP-binding regulatory proteins (G-proteins) by substance P and compound 48/80. FEBS Lett. 1990 Jan 1;259(2):260–262. doi: 10.1016/0014-5793(90)80023-c. [DOI] [PubMed] [Google Scholar]
  24. Nishizuka Y. Studies and perspectives of protein kinase C. Science. 1986 Jul 18;233(4761):305–312. doi: 10.1126/science.3014651. [DOI] [PubMed] [Google Scholar]
  25. Nånberg E., Rozengurt E. Temporal relationship between inositol polyphosphate formation and increases in cytosolic Ca2+ in quiescent 3T3 cells stimulated by platelet-derived growth factor, bombesin and vasopressin. EMBO J. 1988 Sep;7(9):2741–2747. doi: 10.1002/j.1460-2075.1988.tb03128.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Okano Y., Takagi H., Tohmatsu T., Nakashima S., Kuroda Y., Saito K., Nozawa Y. A wasp venom mastoparan-induced polyphosphoinositide breakdown in rat peritoneal mast cells. FEBS Lett. 1985 Sep 2;188(2):363–366. doi: 10.1016/0014-5793(85)80403-8. [DOI] [PubMed] [Google Scholar]
  27. Perianin A., Snyderman R. Mastoparan, a wasp venom peptide, identifies two discrete mechanisms for elevating cytosolic calcium and inositol trisphosphates in human polymorphonuclear leukocytes. J Immunol. 1989 Sep 1;143(5):1669–1673. [PubMed] [Google Scholar]
  28. 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]
  29. Rodriguez-Pena A., Rozengurt E. Phosphorylation of an acidic mol. wt. 80 000 cellular protein in a cell-free system and intact Swiss 3T3 cells: a specific marker of protein kinase C activity. EMBO J. 1986 Jan;5(1):77–83. doi: 10.1002/j.1460-2075.1986.tb04180.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Rodriguez-Pena A., Rozengurt E. Vasopressin rapidly stimulates protein kinase C in quiescent Swiss 3T3 cells. J Cell Physiol. 1986 Oct;129(1):124–130. doi: 10.1002/jcp.1041290117. [DOI] [PubMed] [Google Scholar]
  31. Rozengurt E. Early signals in the mitogenic response. Science. 1986 Oct 10;234(4773):161–166. doi: 10.1126/science.3018928. [DOI] [PubMed] [Google Scholar]
  32. Rozengurt E., Legg A., Pettican P. Vasopressin stimulation of mouse 3T3 cell growth. Proc Natl Acad Sci U S A. 1979 Mar;76(3):1284–1287. doi: 10.1073/pnas.76.3.1284. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Rozengurt E., Murray M., Zachary I., Collins M. Protein kinase C activation enhances cAMP accumulation in Swiss 3T3 cells: inhibition by pertussis toxin. Proc Natl Acad Sci U S A. 1987 Apr;84(8):2282–2286. doi: 10.1073/pnas.84.8.2282. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Rozengurt E., Rodriguez-Pena A., Coombs M., Sinnett-Smith J. Diacylglycerol stimulates DNA synthesis and cell division in mouse 3T3 cells: role of Ca2+-sensitive phospholipid-dependent protein kinase. Proc Natl Acad Sci U S A. 1984 Sep;81(18):5748–5752. doi: 10.1073/pnas.81.18.5748. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Rozengurt E., Rodriguez-Pena M., Smith K. A. Phorbol esters, phospholipase C, and growth factors rapidly stimulate the phosphorylation of a Mr 80,000 protein in intact quiescent 3T3 cells. Proc Natl Acad Sci U S A. 1983 Dec;80(23):7244–7248. doi: 10.1073/pnas.80.23.7244. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Rozengurt E., Sinnett-Smith J. Bombesin stimulation of DNA synthesis and cell division in cultures of Swiss 3T3 cells. Proc Natl Acad Sci U S A. 1983 May;80(10):2936–2940. doi: 10.1073/pnas.80.10.2936. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Rozengurt E., Stroobant P., Waterfield M. D., Deuel T. F., Keehan M. Platelet-derived growth factor elicits cyclic AMP accumulation in Swiss 3T3 cells: role of prostaglandin production. Cell. 1983 Aug;34(1):265–272. doi: 10.1016/0092-8674(83)90157-5. [DOI] [PubMed] [Google Scholar]
  38. Saito H., Okajima F., Molski T. F., Sha'afi R. I., Ui M., Ishizaka T. Effects of ADP-ribosylation of GTP-binding protein by pertussis toxin on immunoglobulin E-dependent and -independent histamine release from mast cells and basophils. J Immunol. 1987 Jun 1;138(11):3927–3934. [PubMed] [Google Scholar]
  39. Tohkin M., Yagami T., Matsubara T. Mastoparan, a peptide toxin from wasp venom, stimulates glycogenolysis mediated by an increase of the cytosolic free Ca2+ concentration but not by an increase of cAMP in rat hepatocytes. FEBS Lett. 1990 Jan 29;260(2):179–182. doi: 10.1016/0014-5793(90)80098-4. [DOI] [PubMed] [Google Scholar]
  40. Weingarten R., Ransnäs L., Mueller H., Sklar L. A., Bokoch G. M. Mastoparan interacts with the carboxyl terminus of the alpha subunit of Gi. J Biol Chem. 1990 Jul 5;265(19):11044–11049. [PubMed] [Google Scholar]
  41. Yokokawa N., Komatsu M., Takeda T., Aizawa T., Yamada T. Mastoparan, a wasp venom, stimulates insulin release by pancreatic islets through pertussis toxin sensitive GTP-binding protein. Biochem Biophys Res Commun. 1989 Feb 15;158(3):712–716. doi: 10.1016/0006-291x(89)92779-4. [DOI] [PubMed] [Google Scholar]
  42. Zachary I., Millar J., Nånberg E., Higgins T., Rozengurt E. Inhibition of bombesin-induced mitogenesis by pertussis toxin: dissociation from phospholipase C pathway. Biochem Biophys Res Commun. 1987 Jul 31;146(2):456–463. doi: 10.1016/0006-291x(87)90551-1. [DOI] [PubMed] [Google Scholar]
  43. Zachary I., Sinnett-Smith J. W., Rozengurt E. Early events elicited by bombesin and structurally related peptides in quiescent Swiss 3T3 cells. I. Activation of protein kinase C and inhibition of epidermal growth factor binding. J Cell Biol. 1986 Jun;102(6):2211–2222. doi: 10.1083/jcb.102.6.2211. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from The Journal of Cell Biology are provided here courtesy of The Rockefeller University Press

RESOURCES