Characterization of hematopoietic lineage-specific gene expression by ES cell in vitro differentiation induction system.
The continuous generation of mature blood cells from hematopoietic progenitor cells requires a highly complex series of molecular events. To examine lineage-specific gene expression during the differentiation process, we developed a novel method combining LacZ reporter gene analysis with in vitro hematopoietic differentiation induction from mouse embryonic stem cells. For a model system using this method, we chose the erythroid and megakaryocytic differentiation pathways. Although erythroid and megakaryocytic cells possess distinct functional and morphologic features, these 2 lineages originate from bipotential erythro-megakaryocytic progenitors and share common lineage-restricted transcription factors. A portion of the 5' flanking region of the human glycoprotein IIb (alphaIIb) integrin gene extending from base -598 to base +33 was examined in detail. As reported previously, this region is sufficient for megakaryocyte-specific gene expression. However, previous reports that used human erythro-megakaryocytic cell lines suggested that one or more negative regulatory regions were necessary for megakaryocyte-specific gene expression. Our data clearly showed that an approximately 200-base enhancer region extending from -598 to -400 was sufficient for megakaryocyte-specific gene expression. This experimental system has advantages over those using erythro-megakaryocytic cell lines because it recapitulates normal hematopoietic cell development and differentiation. Furthermore, this system is more efficient than transgenic analysis and can easily examine gene expression with null mutations of specific genes.
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Department of Molecular Cell Biology, Research Institute for Microbial Diseases, Osaka University, Osaka, Japan.
Takagi
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Takahashi
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Bories
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JC
Nakano
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D000818:Animals; D001665:Binding Sites; D002454:Cell Differentiation / Q000187:drug effects; D019070:Cell Lineage / Q000235:genetics*; D004268:DNA-Binding Proteins / Q000378:metabolism; D004742:Enhancer Elements, Genetic; D004912:Erythrocytes / Q000166:cytology; D015672:Erythroid Precursor Cells / Q000166:cytology / Q000187:drug effects; D050981:Erythroid-Specific DNA-Binding Factors; D004920:Erythropoiesis / Q000187:drug effects; D004921:Erythropoietin / Q000494:pharmacology; D005786:Gene Expression Regulation / Q000187:drug effects*; D005813:Genes, Synthetic; D006410:Hematopoiesis / Q000187:drug effects*; D006801:Humans; D007763:Lac Operon; D016173:Macrophage Colony-Stimulating Factor / Q000494:pharmacology; D008264:Macrophages / Q000166:cytology; D008533:Megakaryocytes / Q000166:cytology / Q000187:drug effects; D051379:Mice; D020648:Peptide Elongation Factor 1 / Q000235:genetics; D019039:Platelet Glycoprotein GPIIb-IIIa Complex / Q000235:genetics; D011518:Proto-Oncogene Proteins / Q000378:metabolism; D050783:Proto-Oncogene Proteins c-ets; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000235:genetics; D012045:Regulatory Sequences, Nucleic Acid; D013234:Stem Cells / Q000166:cytology / Q000187:drug effects*; D013926:Thrombopoietin / Q000494:pharmacology; D014157:Transcription Factors / Q000378:metabolism
D004268:DNA-Binding Proteins; D050981:Erythroid-Specific DNA-Binding Factors; D020648:Peptide Elongation Factor 1; D019039:Platelet Glycoprotein GPIIb-IIIa Complex; D011518:Proto-Oncogene Proteins; D050783:Proto-Oncogene Proteins c-ets; D011993:Recombinant Fusion Proteins; D014157:Transcription Factors; D004921:Erythropoietin; D016173:Macrophage Colony-Stimulating Factor; D013926:Thrombopoietin