Forced GATA-1 expression in the murine myeloid cell line M1: Induction of c-Mpl expression and megakaryocytic/erythroid differentiation

Yuji Yamaguehi, Leonard I. Zon, Steven J. Ackerman, Masayuki Yamamoto, Toshio Suda

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Abstract

The 'zinc-finger' transcription factor GATA-1 was first shown in cells of erythroid lineage. It is also expressed in cells of other hematopoietic lineages including megakaryocytes, mast cells, and eosinophils. GATA-1 is now considered to be one of the central regulators in hematopoietic cell differentiation. To further analyze the role of GATA-1 in controlling differentiation from hematopoietic stem cells, we investigated the phenotypic changes induced by the overexpression of murine GATA-1 in the murine myeloid leukemic cell line, M1. Forced expression of GATA-1 induced the appearance of erythroid cells and megakaryocytes as assessed by cellular morphology, acetylcholinesterase activity, and expression of platelet factor 4 and β- globin mRNA synthesis. Because the c-mpl ligand, thrombopoietin, plays an important role in megakaryopoiesis, the expression of c-mpl and c-mpl ligand (thrombopoietin) mRNA was analyzed by Northern blot and reverse transcription-polymerase chain reaction (RT-PCR) in M1 cells overexpressing GATA-1. The c-mpl ligand mRNA was equally expressed both in parental M1 cells and in those transfected with the GATA-1 expression vector. In contrast, the mRNA expression of c-mpl was increased only in GATA-1 expressing M1 cells differentiated towards erythroid and megakaryocyte lineages. The increased expression of c-mpl mRNA induced by GATA-1 raised the question as to whether or not GATA-1 transactivated the c-mpl promoter. The activity of the c-mpl promoter in the presence of cotransfected GATA-1 was significantly increased compared with that of the control. A plasmid with the mutated GATA-binding site did not show transactivation ability in the cotransfection with a GATA expression vector. These findings suggest that the upregulation of c-mpl induced by GATA-1 expression in M1 cells is closely associated with erythroid and megakaryocytic differentiation.

Original languageEnglish
Pages (from-to)450-457
Number of pages8
JournalBlood
Volume91
Issue number2
Publication statusPublished - 1998 Jan 15

Fingerprint

Thrombopoietin
Myeloid Cells
Cells
Megakaryocytes
Cell Line
Messenger RNA
GATA Transcription Factors
Platelet Factor 4
Erythroid Cells
Globins
Polymerase chain reaction
Zinc Fingers
Cell Lineage
Transcription
Acetylcholinesterase
Hematopoietic Stem Cells
Stem cells
Eosinophils
Mast Cells
Northern Blotting

ASJC Scopus subject areas

  • Hematology

Cite this

Yamaguehi, Y., Zon, L. I., Ackerman, S. J., Yamamoto, M., & Suda, T. (1998). Forced GATA-1 expression in the murine myeloid cell line M1: Induction of c-Mpl expression and megakaryocytic/erythroid differentiation. Blood, 91(2), 450-457.

Forced GATA-1 expression in the murine myeloid cell line M1 : Induction of c-Mpl expression and megakaryocytic/erythroid differentiation. / Yamaguehi, Yuji; Zon, Leonard I.; Ackerman, Steven J.; Yamamoto, Masayuki; Suda, Toshio.

In: Blood, Vol. 91, No. 2, 15.01.1998, p. 450-457.

Research output: Contribution to journalArticle

Yamaguehi, Y, Zon, LI, Ackerman, SJ, Yamamoto, M & Suda, T 1998, 'Forced GATA-1 expression in the murine myeloid cell line M1: Induction of c-Mpl expression and megakaryocytic/erythroid differentiation', Blood, vol. 91, no. 2, pp. 450-457.
Yamaguehi, Yuji ; Zon, Leonard I. ; Ackerman, Steven J. ; Yamamoto, Masayuki ; Suda, Toshio. / Forced GATA-1 expression in the murine myeloid cell line M1 : Induction of c-Mpl expression and megakaryocytic/erythroid differentiation. In: Blood. 1998 ; Vol. 91, No. 2. pp. 450-457.
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abstract = "The 'zinc-finger' transcription factor GATA-1 was first shown in cells of erythroid lineage. It is also expressed in cells of other hematopoietic lineages including megakaryocytes, mast cells, and eosinophils. GATA-1 is now considered to be one of the central regulators in hematopoietic cell differentiation. To further analyze the role of GATA-1 in controlling differentiation from hematopoietic stem cells, we investigated the phenotypic changes induced by the overexpression of murine GATA-1 in the murine myeloid leukemic cell line, M1. Forced expression of GATA-1 induced the appearance of erythroid cells and megakaryocytes as assessed by cellular morphology, acetylcholinesterase activity, and expression of platelet factor 4 and β- globin mRNA synthesis. Because the c-mpl ligand, thrombopoietin, plays an important role in megakaryopoiesis, the expression of c-mpl and c-mpl ligand (thrombopoietin) mRNA was analyzed by Northern blot and reverse transcription-polymerase chain reaction (RT-PCR) in M1 cells overexpressing GATA-1. The c-mpl ligand mRNA was equally expressed both in parental M1 cells and in those transfected with the GATA-1 expression vector. In contrast, the mRNA expression of c-mpl was increased only in GATA-1 expressing M1 cells differentiated towards erythroid and megakaryocyte lineages. The increased expression of c-mpl mRNA induced by GATA-1 raised the question as to whether or not GATA-1 transactivated the c-mpl promoter. The activity of the c-mpl promoter in the presence of cotransfected GATA-1 was significantly increased compared with that of the control. A plasmid with the mutated GATA-binding site did not show transactivation ability in the cotransfection with a GATA expression vector. These findings suggest that the upregulation of c-mpl induced by GATA-1 expression in M1 cells is closely associated with erythroid and megakaryocytic differentiation.",
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