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. 1996 May 15;316(Pt 1):93–98. doi: 10.1042/bj3160093

Thrombopoietin potentiates activation of human platelets in association with JAK2 and TYK2 phosphorylation.

B Rodríguez-Liñares 1, S P Watson 1
PMCID: PMC1217355  PMID: 8645238

Abstract

Thrombopoietin (TPO), also known as the c-mpl ligand, stimulates rapid tyrosine phosphorylation of multiple proteins in human platelets including the Janus family kinases JAK2 and TYK2. On its own, TPO has no effect on platelet aggregation and dense-granule secretion but induces a general potentiation of these responses by other stimuli. The most dramatic effect is observed against threshold concentrations of agonists for aggregation. Shape change or weak reversible aggregation induced by low concentrations of thrombin, collagen and the thromboxane mimetic, U46619, are converted into irreversible aggregation in the presence of TPO. A similar result is obtained in the presence of the ADP scavenger apyrase and cyclo-oxygenase inhibitor indomethacin. TPO also induces potentiation of dense-granule secretion measured through release of 5-hydroxy[3H]-tryptamine. This effect is most striking against low concentrations of stimuli and is independent of aggregation as it is observed in the presence of chelation of extracellular Ca2+ with EGTA. TPO potentiates activation of phospholipase C and elevation of intracellular Ca2+, providing a molecular explanation for potentiation of functional responses. TPO may have an important physiological role in priming platelet activation in thrombocytopenia, an action that may help to compensate for the reduced platelet density.

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Selected References

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  1. Argetsinger L. S., Campbell G. S., Yang X., Witthuhn B. A., Silvennoinen O., Ihle J. N., Carter-Su C. Identification of JAK2 as a growth hormone receptor-associated tyrosine kinase. Cell. 1993 Jul 30;74(2):237–244. doi: 10.1016/0092-8674(93)90415-m. [DOI] [PubMed] [Google Scholar]
  2. Bartley T. D., Bogenberger J., Hunt P., Li Y. S., Lu H. S., Martin F., Chang M. S., Samal B., Nichol J. L., Swift S. Identification and cloning of a megakaryocyte growth and development factor that is a ligand for the cytokine receptor Mpl. Cell. 1994 Jul 1;77(7):1117–1124. doi: 10.1016/0092-8674(94)90450-2. [DOI] [PubMed] [Google Scholar]
  3. Choi E. S., Nichol J. L., Hokom M. M., Hornkohl A. C., Hunt P. Platelets generated in vitro from proplatelet-displaying human megakaryocytes are functional. Blood. 1995 Jan 15;85(2):402–413. [PubMed] [Google Scholar]
  4. Coorssen J. R., Haslam R. J. GTP gamma S and phorbol ester act synergistically to stimulate both Ca(2+)-independent secretion and phospholipase D activity in permeabilized human platelets. Inhibition by BAPTA and analogues. FEBS Lett. 1993 Jan 25;316(2):170–174. doi: 10.1016/0014-5793(93)81209-i. [DOI] [PubMed] [Google Scholar]
  5. Debili N., Wendling F., Cosman D., Titeux M., Florindo C., Dusanter-Fourt I., Schooley K., Methia N., Charon M., Nador R. The Mpl receptor is expressed in the megakaryocytic lineage from late progenitors to platelets. Blood. 1995 Jan 15;85(2):391–401. [PubMed] [Google Scholar]
  6. Drachman J. G., Griffin J. D., Kaushansky K. The c-Mpl ligand (thrombopoietin) stimulates tyrosine phosphorylation of Jak2, Shc, and c-Mpl. J Biol Chem. 1995 Mar 10;270(10):4979–4982. doi: 10.1074/jbc.270.10.4979. [DOI] [PubMed] [Google Scholar]
  7. Farese A. M., Hunt P., Boone T., MacVittie T. J. Recombinant human megakaryocyte growth and development factor stimulates thrombocytopoiesis in normal nonhuman primates. Blood. 1995 Jul 1;86(1):54–59. [PubMed] [Google Scholar]
  8. Fukunaga R., Ishizaka-Ikeda E., Nagata S. Purification and characterization of the receptor for murine granulocyte colony-stimulating factor. J Biol Chem. 1990 Aug 15;265(23):14008–14015. [PubMed] [Google Scholar]
  9. Grabarek J., Groopman J. E., Lyles Y. R., Jiang S., Bennett L., Zsebo K., Avraham H. Human kit ligand (stem cell factor) modulates platelet activation in vitro. J Biol Chem. 1994 Aug 26;269(34):21718–21724. [PubMed] [Google Scholar]
  10. Gurney A. L., Wong S. C., Henzel W. J., de Sauvage F. J. Distinct regions of c-Mpl cytoplasmic domain are coupled to the JAK-STAT signal transduction pathway and Shc phosphorylation. Proc Natl Acad Sci U S A. 1995 Jun 6;92(12):5292–5296. doi: 10.1073/pnas.92.12.5292. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Ihle J. N., Kerr I. M. Jaks and Stats in signaling by the cytokine receptor superfamily. Trends Genet. 1995 Feb;11(2):69–74. doi: 10.1016/s0168-9525(00)89000-9. [DOI] [PubMed] [Google Scholar]
  12. Ihle J. N., Witthuhn B. A., Quelle F. W., Yamamoto K., Thierfelder W. E., Kreider B., Silvennoinen O. Signaling by the cytokine receptor superfamily: JAKs and STATs. Trends Biochem Sci. 1994 May;19(5):222–227. doi: 10.1016/0968-0004(94)90026-4. [DOI] [PubMed] [Google Scholar]
  13. Kaushansky K., Broudy V. C., Lin N., Jorgensen M. J., McCarty J., Fox N., Zucker-Franklin D., Lofton-Day C. Thrombopoietin, the Mp1 ligand, is essential for full megakaryocyte development. Proc Natl Acad Sci U S A. 1995 Apr 11;92(8):3234–3238. doi: 10.1073/pnas.92.8.3234. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Kaushansky K., Lok S., Holly R. D., Broudy V. C., Lin N., Bailey M. C., Forstrom J. W., Buddle M. M., Oort P. J., Hagen F. S. Promotion of megakaryocyte progenitor expansion and differentiation by the c-Mpl ligand thrombopoietin. Nature. 1994 Jun 16;369(6481):568–571. doi: 10.1038/369568a0. [DOI] [PubMed] [Google Scholar]
  15. Kaushansky K. Thrombopoietin: the primary regulator of platelet production. Blood. 1995 Jul 15;86(2):419–431. [PubMed] [Google Scholar]
  16. Kuter D. J., Rosenberg R. D. The reciprocal relationship of thrombopoietin (c-Mpl ligand) to changes in the platelet mass during busulfan-induced thrombocytopenia in the rabbit. Blood. 1995 May 15;85(10):2720–2730. [PubMed] [Google Scholar]
  17. Lok S., Kaushansky K., Holly R. D., Kuijper J. L., Lofton-Day C. E., Oort P. J., Grant F. J., Heipel M. D., Burkhead S. K., Kramer J. M. Cloning and expression of murine thrombopoietin cDNA and stimulation of platelet production in vivo. Nature. 1994 Jun 16;369(6481):565–568. doi: 10.1038/369565a0. [DOI] [PubMed] [Google Scholar]
  18. Methia N., Louache F., Vainchenker W., Wendling F. Oligodeoxynucleotides antisense to the proto-oncogene c-mpl specifically inhibit in vitro megakaryocytopoiesis. Blood. 1993 Sep 1;82(5):1395–1401. [PubMed] [Google Scholar]
  19. Miyakawa Y., Oda A., Druker B. J., Kato T., Miyazaki H., Handa M., Ikeda Y. Recombinant thrombopoietin induces rapid protein tyrosine phosphorylation of Janus kinase 2 and Shc in human blood platelets. Blood. 1995 Jul 1;86(1):23–27. [PubMed] [Google Scholar]
  20. Nunn D. L., Watson S. P. A diacylglycerol kinase inhibitor, R59022, potentiates secretion by and aggregation of thrombin-stimulated human platelets. Biochem J. 1987 May 1;243(3):809–813. doi: 10.1042/bj2430809. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Poenie M. Alteration of intracellular Fura-2 fluorescence by viscosity: a simple correction. Cell Calcium. 1990 Feb-Mar;11(2-3):85–91. doi: 10.1016/0143-4160(90)90062-y. [DOI] [PubMed] [Google Scholar]
  22. Rui H., Kirken R. A., Farrar W. L. Activation of receptor-associated tyrosine kinase JAK2 by prolactin. J Biol Chem. 1994 Feb 18;269(7):5364–5368. [PubMed] [Google Scholar]
  23. Shimoda K., Okamura S., Harada N., Kondo S., Okamura T., Niho Y. Identification of a functional receptor for granulocyte colony-stimulating factor on platelets. J Clin Invest. 1993 Apr;91(4):1310–1313. doi: 10.1172/JCI116330. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Skoda R. C., Seldin D. C., Chiang M. K., Peichel C. L., Vogt T. F., Leder P. Murine c-mpl: a member of the hematopoietic growth factor receptor superfamily that transduces a proliferative signal. EMBO J. 1993 Jul;12(7):2645–2653. doi: 10.1002/j.1460-2075.1993.tb05925.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Souyri M., Vigon I., Penciolelli J. F., Heard J. M., Tambourin P., Wendling F. A putative truncated cytokine receptor gene transduced by the myeloproliferative leukemia virus immortalizes hematopoietic progenitors. Cell. 1990 Dec 21;63(6):1137–1147. doi: 10.1016/0092-8674(90)90410-g. [DOI] [PubMed] [Google Scholar]
  26. Taniguchi T. Cytokine signaling through nonreceptor protein tyrosine kinases. Science. 1995 Apr 14;268(5208):251–255. doi: 10.1126/science.7716517. [DOI] [PubMed] [Google Scholar]
  27. Tortolani P. J., Johnston J. A., Bacon C. M., McVicar D. W., Shimosaka A., Linnekin D., Longo D. L., O'Shea J. J. Thrombopoietin induces tyrosine phosphorylation and activation of the Janus kinase, JAK2. Blood. 1995 Jun 15;85(12):3444–3451. [PubMed] [Google Scholar]
  28. Vigon I., Florindo C., Fichelson S., Guenet J. L., Mattei M. G., Souyri M., Cosman D., Gisselbrecht S. Characterization of the murine Mpl proto-oncogene, a member of the hematopoietic cytokine receptor family: molecular cloning, chromosomal location and evidence for a function in cell growth. Oncogene. 1993 Oct;8(10):2607–2615. [PubMed] [Google Scholar]
  29. Vigon I., Mornon J. P., Cocault L., Mitjavila M. T., Tambourin P., Gisselbrecht S., Souyri M. Molecular cloning and characterization of MPL, the human homolog of the v-mpl oncogene: identification of a member of the hematopoietic growth factor receptor superfamily. Proc Natl Acad Sci U S A. 1992 Jun 15;89(12):5640–5644. doi: 10.1073/pnas.89.12.5640. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Walker T. R., Watson S. P. Synergy between Ca2+ and protein kinase C is the major factor in determining the level of secretion from human platelets. Biochem J. 1993 Jan 1;289(Pt 1):277–282. doi: 10.1042/bj2890277. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Watson S. P., McNally J., Shipman L. J., Godfrey P. P. The action of the protein kinase C inhibitor, staurosporine, on human platelets. Evidence against a regulatory role for protein kinase C in the formation of inositol trisphosphate by thrombin. Biochem J. 1988 Jan 15;249(2):345–350. doi: 10.1042/bj2490345. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Witthuhn B. A., Quelle F. W., Silvennoinen O., Yi T., Tang B., Miura O., Ihle J. N. JAK2 associates with the erythropoietin receptor and is tyrosine phosphorylated and activated following stimulation with erythropoietin. Cell. 1993 Jul 30;74(2):227–236. doi: 10.1016/0092-8674(93)90414-l. [DOI] [PubMed] [Google Scholar]
  33. de Sauvage F. J., Hass P. E., Spencer S. D., Malloy B. E., Gurney A. L., Spencer S. A., Darbonne W. C., Henzel W. J., Wong S. C., Kuang W. J. Stimulation of megakaryocytopoiesis and thrombopoiesis by the c-Mpl ligand. Nature. 1994 Jun 16;369(6481):533–538. doi: 10.1038/369533a0. [DOI] [PubMed] [Google Scholar]

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