Promoter motifs required for c-mpl gene expression induced by thrombopoietin in CMK cells

Masataka Sunohara; Iwao Sato; Shigeru Morikawa

doi:10.14715/cmb/2017.63.11.21

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Vol. 63 No. 11: Issue 11

Issue Published : November 30, 2017

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Promoter motifs required for c-mpl gene expression induced by thrombopoietin in CMK cells

https://doi.org/10.14715/cmb/2017.63.11.21

Masataka Sunohara

Department of Anatomy, School of Life Dentistry at Tokyo, The Nippon Dental University,1-9-20 Fujimi, Chiyoda-ku, Tokyo 102-8159, Japan

Iwao Sato

Department of Anatomy, School of Life Dentistry at Tokyo, The Nippon Dental University,1-9-20 Fujimi, Chiyoda-ku, Tokyo 102-8159, Japan

Shigeru Morikawa

Department of Veterinary Science, National Institute of Infectious Diseases, 1-23-1 Toyama, Shinjuku-ku, Tokyo 162-8640, Japan

Corresponding Author(s) : Masataka Sunohara

ma-suno@tky.ndu.ac.jp

Cellular and Molecular Biology, Vol. 63 No. 11: Issue 11
Article Published : November 30, 2017

Abstract

Thrombopoietin (TPO) and its receptor, c-Mpl, are the central regulators of megakaryocyte development and platelet production and are also crucial to regulate megakaryocytopoiesis. TPO remarkably elevated c-mpl promoter activity, while the protein kinase C (PKC) inhibitors, GF109203, H7 and Calphostin C, clearly reduced the steady level of its promoter activity. In the present study, motifs crucial for c-mpl promoter activity induced by TPO treatment has been analyzed using a human megakaryoblastic cell line, CMK. Destruction of the –107Sp1 and the –57Sp1 sites in the c-mpl promoter enhancer region resulted in decrease of the promoter activity by 53.1% and 64.4%, respectively, and destruction of –69Ets and –28Ets elements dramatically decreased the promoter activity by 96.4% and 87.8%, respectively, while mutation of –77GATA moderately reduced the activity by 31.4%. The result was in agreement with our previous report that showed the crucial motifs in the c-mpl promoter for the promoter activity induced by PMA-treatment. This indicates that TPO-induced activation of the c-mpl promoter activity is fully modulated by transcription through a PKC-dependent pathway and the two Sp1 and two Ets motifs are crucial for the activation of the c-mpl promoter activity rather than a GATA motif in the c-mpl promoter of CMK cells.

Keywords

Thrombopoietin c-mpl Protein kinase C Megakaryocyte CMK.

Sunohara, M., Sato, I., & Morikawa, S. (2017). Promoter motifs required for c-mpl gene expression induced by thrombopoietin in CMK cells. Cellular and Molecular Biology, 63(11), 119–123. https://doi.org/10.14715/cmb/2017.63.11.21

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References

Bartley TD, Bogenberger J, Hunt P, Li YS, Lu HS, Martin F, et al. Identification and cloning of a megakaryocyte growth and development factor that is a ligand for the cytokine receptor Mpl. Cell 1994; 77: 1117-24.
Ellis MH, Avraham H, Groopman JE. The regulation of megakaryocytopoiesis. Blood Rev 1995; 9 : 1-6.
Kaushansky K. Thrombopoietin: the primary regulator of platelet production. Blood 1995; 86: 419-31.
De Sauvage FJ, Hass PE, Spencer SD, Malloy BE, Gurney AL, Spencer SA, et al. Stimulation of megakaryocytopoesis and thrombopoiesis by the c-Mpl ligand. Nature 1994; 369: 533-8.
Lok S, Kaushansky K, Holly RD, Kuijper JL, Lofton-Day CE, Oort PJ, et al. Cloning and expression of murine thrombopoietin cDNA and stimulation of platelet production in vivo. Nature 1994; 369: 565-8.
Wendling F, Maraskovsky E, Debili N, Florindo C, Teepe M, Titeux M, et al. c-Mpl ligand is a humoral regulator of megakaryocytopoiesis. Nature 1994; 369: 571-4.
Kaushansky K. The enigmatic megakaryocyte gradually reveals its secrets. Bioessays 1999; 21: 353-60.
Gurney A, Carver-Moore K, de Sauvage FJ, Moore MW. Thrombopoietin in c-mpl-deficient mice. Science 1994; 265:1445-7.
Alexander WS, Roberts AW, Nicola NA, Li R, Metcalf D. Deficiencies in progenitor cells of multiple hematopoietic lineages and defective megakaryocytopoiesis in mice lacking the thrombopoietic receptor c-Mpl. Blood 1996; 87: 2162-70.
De Sauvage FJ, Carver-Moore K, Luoh SM, Ryan A, Dowd M, Eaton DL, et al. Physiological regulation of early and late stages of megakaryocytopoiesis by thrombopoietin. J Exp Med 1996;183: 651-6.
Yee NS, Hsiau CW, Serve H, Vosseller K, Besmer P. Mechanism of down-regulation of c-kit receptor. Roles of receptor tyrosine kinase, phosphatidylinositol 3'-kinase, and protein kinase C. J Biol Chem 1994; 269 : 31991-8.
Downing JR, Roussel MF, Sher CJ. Ligand and protein kinase C downmodulate the colony-stimulation factor 1 receptor by independent mechanisms. Mol Cell Biol 1989; 9 :2890-6.
Porteu F, Nathan C. Shedding of tumor necrosis factor receptors by activated human neutrophils. J Exp Med 1990; 172: 599-607.
Yee NS, Langen H, Besmer P. Mechanism of kit ligand, phorbol ester, and calcium-induced down-regulation of c-kit receptors in mast cells. J Biol Chem 1993; 268: 14189-201.
Yang Z, Huerlimann DA, Viswambharan H, Luescher TF.
Phorbol ester downregulates PDGFbeta receptor via PKCbeta1 in vascular
smooth muscle cells. Biochem Biophys Res Commun 2001;286 372-5.
Sato T, Fuse A, Eguchi M, Hayashi Y, Ryo R, Adachi M, et al. Establishment of a human leukaemic cell line (CMK) with magakaryocytic characteristics from a Down's syndrome patient with acute megakaryoblastic leukemia. Br. J. Haematol 1989; 72: 184-90.
Sato T, Fuse A, Niimi H, Fielder PJ, Avraham H. Binding and regulation of thrombopoietin to human megakaryocytes. Br J Haematol 1998; 100: 704-11.
Millot GA, Vainchenker W, Dumenil D, Svinarchuk F. Distinct effects of thrombopoietin depending on a threshold level of activated Mpl in BaF-3 cells. J Cell Sci 2002; 115(Pt11): 2329-37.
Hong Y, Dumenil D, van der Loo B, Goncalves F, Vainchenker W, Erusalimsky JD. Protein kinase C mediates the mitogenic action of thrombopoietin in c-Mpl-expressing UT-7 cells. Blood. 1998; 91(3):813-22.
Sunohara M, Morikawa S, Sato T, Sato I, Sato T, Fuse A. Modulation of human c-mpl gene expression by thrombopoietin through protein kinase C. Cell Mol Biol 2003; 49: Online Pub: OL393-8.
Ohno S, Akita Y, Hata A, Osada S, Kubo K, Konno Y, et al. Structural and functional diversities of a family of signal transducing protein kinases, protein kinase C family: two distinct classes of PKC, conventional cPKC and novel nPKC. Adv Enzyme Regul 1991; 31: 287-303.
Nishizuka Y. Intracellular signaling by hydrolysis of phospholipids and activation of protein kinase C. Science 1992; 258: 607-14.
Nagata KI, Okano Y, Nozawa Y. Protein kinase C isozymes in human megakaryoblastic leukemia cell line, MEG-01: possible involvement of the isozymes in the differentiation process of MEG-01 cells. Br J Haematol 1996; 93:762-71.
Thabard W, Collette M, Bataille R, Amiot M. Protein kinase C delta and eta isoenzymes control the shedding of the interleukin 6 receptor alpha in myeloma cells. Biochem J 2001; 358: 193-200.
Komatsu N, Suda T, Moroi M, Tokuyama N, Sakata Y, Okada M, et al. Growth and differentiation of a human megakaryoblastic cell line CMK. Blood 1989; 74: 42-8.
Sunohara M, Morikawa S, Sato T, Miyado M, Sato I, Sato T, Fuse A.
Promoter regulatory motifs involved in c-mpl gene expression induced by PMA.
Cell Biol Int. 2008; 32:692-7.
Prandini MH, Uzan G, Martin F, Thevenon D, Marguerie G.
Characterrization of a specific erythromegakaryocytic enhancer within the glycoprotein IIb promoter. J Biol Chem 1992; 267:10370-4.
Yamaguchi Y, Zon LI, Ackerman SJ, Yamamoto M, Suda T. Forced GATA-1 expression in the murine myeloid cell line M1: induction of c-Mpl expression and megakaryocytic/erythroid differentiation. Blood 1998; 91: 450-7.
Vyas P, Ault K, Jackson CW, Orkin SH, Shivdasani RA.
Consequences of GATA-1 Deficiency in Megakaryocytes and Platelets.
Blood 1999; 93 : 2867-75.
Sunohara M, Morikawa S, Fuse A, Sato I. Role of promoter element in c-mpl gene expression induced by TPO. Okajimas Folia Anat Jpn. 2013;89:131-5.
Parganas E, Wang D, Stravopodis D, Topham DJ, Marine JC, Teglund S, et al.
Jak2 is essential for signaling through a variety of cytokine receptors.
Cell. 1998;93: 385-395.
Schulze H1, Ballmaier M, Welte K, Germeshausen M. Thrombopoietin induces the generation of distinct Stat1, Stat3, Stat5a and Stat5b homo- and heterodimeric complexes with different kinetics in human platelets.
Exp Hematol. 2000;28: 294-304.
Kaushansky K, Drachman JG. The molecular and cellular biology of thrombopoietin: the primary regulator of platelet production.
Oncogene. 2002; 21:3359-67.
Dorsch M, Fan PD, Danial NN, Rothman PB, Goff SP.
The thrombopoietin receptor can mediate proliferation without activation of the Jak-STAT pathway.
J Exp Med. 1997; 186:1947-55.
Lemarchandel V, Ghysdael J, Mignotte V, Rahuel C, Romeo PH. GATA and Ets cis-acting sequences mediate megakaryocyte-specific expression. Mol Cell Biol 1993; 13: 668-76.
Ogami K. Gene expression and transcriptional regulation of thrombopoietin. Stem Cells 1996; 14 :148-53.
Kamura T, Hanada H, Hamasaki N, Kitajima S. Characterzation of the human thrombopoietin gene promoter: a possible role of an Ets transcription factor, E4TF1/GABP. J Biol Chem 1997; 272:11361-8.
D'Angelo DD, Oliver BG, Davis MG, McCluskey TS, Dorn GW. Novel role for Sp1 in phorbol ester enhancement of human platelet thromboxane receptor gene expression. J Biol Chem 1996; 271:19696-704.
Deveaux S, Filipe A, Lemarchandel V, Ghysdael J, Romeo PH, Mignotte V. Analysis of the thrombopoietin receptor (MPL) promoter implicates GATA and Ets proteins in the coregulation of megakaryocyte-specific genes. Blood. 1996;87(11):4678-85.
You HL, Eng HL, Hsu SF, Chen CM, Ye TC, Liao WT, Huang MY, Baer R, Cheng JT. A PKC-Sp1 signaling pathway induces early differentiation of human keratinocytes through upregulation of TSG101. Cell Signal. 2007; 19(6):1201-11.

References

Bartley TD, Bogenberger J, Hunt P, Li YS, Lu HS, Martin F, et al. Identification and cloning of a megakaryocyte growth and development factor that is a ligand for the cytokine receptor Mpl. Cell 1994; 77: 1117-24.

Ellis MH, Avraham H, Groopman JE. The regulation of megakaryocytopoiesis. Blood Rev 1995; 9 : 1-6.

Kaushansky K. Thrombopoietin: the primary regulator of platelet production. Blood 1995; 86: 419-31.

De Sauvage FJ, Hass PE, Spencer SD, Malloy BE, Gurney AL, Spencer SA, et al. Stimulation of megakaryocytopoesis and thrombopoiesis by the c-Mpl ligand. Nature 1994; 369: 533-8.

Lok S, Kaushansky K, Holly RD, Kuijper JL, Lofton-Day CE, Oort PJ, et al. Cloning and expression of murine thrombopoietin cDNA and stimulation of platelet production in vivo. Nature 1994; 369: 565-8.

Wendling F, Maraskovsky E, Debili N, Florindo C, Teepe M, Titeux M, et al. c-Mpl ligand is a humoral regulator of megakaryocytopoiesis. Nature 1994; 369: 571-4.

Kaushansky K. The enigmatic megakaryocyte gradually reveals its secrets. Bioessays 1999; 21: 353-60.

Gurney A, Carver-Moore K, de Sauvage FJ, Moore MW. Thrombopoietin in c-mpl-deficient mice. Science 1994; 265:1445-7.

Alexander WS, Roberts AW, Nicola NA, Li R, Metcalf D. Deficiencies in progenitor cells of multiple hematopoietic lineages and defective megakaryocytopoiesis in mice lacking the thrombopoietic receptor c-Mpl. Blood 1996; 87: 2162-70.

De Sauvage FJ, Carver-Moore K, Luoh SM, Ryan A, Dowd M, Eaton DL, et al. Physiological regulation of early and late stages of megakaryocytopoiesis by thrombopoietin. J Exp Med 1996;183: 651-6.

Yee NS, Hsiau CW, Serve H, Vosseller K, Besmer P. Mechanism of down-regulation of c-kit receptor. Roles of receptor tyrosine kinase, phosphatidylinositol 3'-kinase, and protein kinase C. J Biol Chem 1994; 269 : 31991-8.

Downing JR, Roussel MF, Sher CJ. Ligand and protein kinase C downmodulate the colony-stimulation factor 1 receptor by independent mechanisms. Mol Cell Biol 1989; 9 :2890-6.

Porteu F, Nathan C. Shedding of tumor necrosis factor receptors by activated human neutrophils. J Exp Med 1990; 172: 599-607.

Yee NS, Langen H, Besmer P. Mechanism of kit ligand, phorbol ester, and calcium-induced down-regulation of c-kit receptors in mast cells. J Biol Chem 1993; 268: 14189-201.

Yang Z, Huerlimann DA, Viswambharan H, Luescher TF.

Phorbol ester downregulates PDGFbeta receptor via PKCbeta1 in vascular

smooth muscle cells. Biochem Biophys Res Commun 2001;286 372-5.

Sato T, Fuse A, Eguchi M, Hayashi Y, Ryo R, Adachi M, et al. Establishment of a human leukaemic cell line (CMK) with magakaryocytic characteristics from a Down's syndrome patient with acute megakaryoblastic leukemia. Br. J. Haematol 1989; 72: 184-90.

Sato T, Fuse A, Niimi H, Fielder PJ, Avraham H. Binding and regulation of thrombopoietin to human megakaryocytes. Br J Haematol 1998; 100: 704-11.

Millot GA, Vainchenker W, Dumenil D, Svinarchuk F. Distinct effects of thrombopoietin depending on a threshold level of activated Mpl in BaF-3 cells. J Cell Sci 2002; 115(Pt11): 2329-37.

Hong Y, Dumenil D, van der Loo B, Goncalves F, Vainchenker W, Erusalimsky JD. Protein kinase C mediates the mitogenic action of thrombopoietin in c-Mpl-expressing UT-7 cells. Blood. 1998; 91(3):813-22.

Sunohara M, Morikawa S, Sato T, Sato I, Sato T, Fuse A. Modulation of human c-mpl gene expression by thrombopoietin through protein kinase C. Cell Mol Biol 2003; 49: Online Pub: OL393-8.

Ohno S, Akita Y, Hata A, Osada S, Kubo K, Konno Y, et al. Structural and functional diversities of a family of signal transducing protein kinases, protein kinase C family: two distinct classes of PKC, conventional cPKC and novel nPKC. Adv Enzyme Regul 1991; 31: 287-303.

Nishizuka Y. Intracellular signaling by hydrolysis of phospholipids and activation of protein kinase C. Science 1992; 258: 607-14.

Nagata KI, Okano Y, Nozawa Y. Protein kinase C isozymes in human megakaryoblastic leukemia cell line, MEG-01: possible involvement of the isozymes in the differentiation process of MEG-01 cells. Br J Haematol 1996; 93:762-71.

Thabard W, Collette M, Bataille R, Amiot M. Protein kinase C delta and eta isoenzymes control the shedding of the interleukin 6 receptor alpha in myeloma cells. Biochem J 2001; 358: 193-200.

Komatsu N, Suda T, Moroi M, Tokuyama N, Sakata Y, Okada M, et al. Growth and differentiation of a human megakaryoblastic cell line CMK. Blood 1989; 74: 42-8.

Sunohara M, Morikawa S, Sato T, Miyado M, Sato I, Sato T, Fuse A.

Promoter regulatory motifs involved in c-mpl gene expression induced by PMA.

Cell Biol Int. 2008; 32:692-7.

Prandini MH, Uzan G, Martin F, Thevenon D, Marguerie G.

Characterrization of a specific erythromegakaryocytic enhancer within the glycoprotein IIb promoter. J Biol Chem 1992; 267:10370-4.

Yamaguchi Y, Zon LI, Ackerman SJ, Yamamoto M, Suda T. Forced GATA-1 expression in the murine myeloid cell line M1: induction of c-Mpl expression and megakaryocytic/erythroid differentiation. Blood 1998; 91: 450-7.

Vyas P, Ault K, Jackson CW, Orkin SH, Shivdasani RA.

Consequences of GATA-1 Deficiency in Megakaryocytes and Platelets.

Blood 1999; 93 : 2867-75.

Sunohara M, Morikawa S, Fuse A, Sato I. Role of promoter element in c-mpl gene expression induced by TPO. Okajimas Folia Anat Jpn. 2013;89:131-5.

Parganas E, Wang D, Stravopodis D, Topham DJ, Marine JC, Teglund S, et al.

Jak2 is essential for signaling through a variety of cytokine receptors.

Cell. 1998;93: 385-395.

Schulze H1, Ballmaier M, Welte K, Germeshausen M. Thrombopoietin induces the generation of distinct Stat1, Stat3, Stat5a and Stat5b homo- and heterodimeric complexes with different kinetics in human platelets.

Exp Hematol. 2000;28: 294-304.

Kaushansky K, Drachman JG. The molecular and cellular biology of thrombopoietin: the primary regulator of platelet production.

Oncogene. 2002; 21:3359-67.

Dorsch M, Fan PD, Danial NN, Rothman PB, Goff SP.

The thrombopoietin receptor can mediate proliferation without activation of the Jak-STAT pathway.

J Exp Med. 1997; 186:1947-55.

Lemarchandel V, Ghysdael J, Mignotte V, Rahuel C, Romeo PH. GATA and Ets cis-acting sequences mediate megakaryocyte-specific expression. Mol Cell Biol 1993; 13: 668-76.

Ogami K. Gene expression and transcriptional regulation of thrombopoietin. Stem Cells 1996; 14 :148-53.

Kamura T, Hanada H, Hamasaki N, Kitajima S. Characterzation of the human thrombopoietin gene promoter: a possible role of an Ets transcription factor, E4TF1/GABP. J Biol Chem 1997; 272:11361-8.

D'Angelo DD, Oliver BG, Davis MG, McCluskey TS, Dorn GW. Novel role for Sp1 in phorbol ester enhancement of human platelet thromboxane receptor gene expression. J Biol Chem 1996; 271:19696-704.

Deveaux S, Filipe A, Lemarchandel V, Ghysdael J, Romeo PH, Mignotte V. Analysis of the thrombopoietin receptor (MPL) promoter implicates GATA and Ets proteins in the coregulation of megakaryocyte-specific genes. Blood. 1996;87(11):4678-85.

You HL, Eng HL, Hsu SF, Chen CM, Ye TC, Liao WT, Huang MY, Baer R, Cheng JT. A PKC-Sp1 signaling pathway induces early differentiation of human keratinocytes through upregulation of TSG101. Cell Signal. 2007; 19(6):1201-11.

Author biographies is not available.