Characterization of the promoter controlling Mona/Gads expression in the megakaryocytic lineage.
Mona/grb2 related adapter downstream of shc is a molecular adapter expressed in platelets, T lymphocytes and myelomonocytic cells. Using human hematopoietic cell lines, we have previously shown that lineage-specific Mona expression is achieved through the production of two transcripts (named 1A and 1B) differing by their 5' untranslated region (5'UTR). Thus, platelets and megakaryocytic cell lines K562 and HEL (Human Erythro-Leukemia) specifically express 1B messenger RNA (mRNA). We report here characterization of the (-2031/+72) genomic region relative to the putative transcription start site of 1B mRNA. We show this region is sufficient to ensure specific reporter gene expression in megakaryocytic cell lines, and that most promoter activity is contained in the (-225/+72) fragment. Electro-mobility shift assay and mutational analyses indicated that GATA-1 and a yet unidentified E-26 family member transcription factor are required for 1B (-2031/+72) promoter activity. Thus, Mona 1B promoter exhibits typical features of megakaryocyte-specific promoters.
Guyot
Boris
B
Centre de Génétique Moléculaire et Cellulaire, UMR CNRS 5534, Université Claude Bernard Lyon-1, 43, boulevard du 11 novembre 1918, Bâtiment Gregor Mendel, 16 rue Raphael Dubois, 69622 Villeurbanne Cedex, France.
Mouchiroud
Guy
G
D048868:Adaptor Proteins, Signal Transducing; D001483:Base Sequence; D001665:Binding Sites / Q000235:genetics; D002352:Carrier Proteins / Q000235:genetics*; D002454:Cell Differentiation / Q000235:genetics; D004268:DNA-Binding Proteins / Q000378:metabolism; D024202:Electrophoretic Mobility Shift Assay; D050981:Erythroid-Specific DNA-Binding Factors; D050982:GATA1 Transcription Factor; D005786:Gene Expression Regulation / Q000187:drug effects; D006801:Humans; D019169:Jurkat Cells; D020014:K562 Cells; D008156:Luciferases / Q000235:genetics / Q000378:metabolism; D008533:Megakaryocytes / Q000378:metabolism* / Q000473:pathology; D008969:Molecular Sequence Data; D011401:Promoter Regions, Genetic / Q000235:genetics*; D011485:Protein Binding; D011518:Proto-Oncogene Proteins / Q000378:metabolism; D050783:Proto-Oncogene Proteins c-ets; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D012045:Regulatory Sequences, Nucleic Acid / Q000235:genetics; D017384:Sequence Deletion; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D014157:Transcription Factors / Q000378:metabolism; D014162:Transfection; D014407:Tumor Cells, Cultured; D020298:U937 Cells; D001616:beta-Galactosidase / Q000235:genetics / Q000378:metabolism
D048868:Adaptor Proteins, Signal Transducing; D002352:Carrier Proteins; D004268:DNA-Binding Proteins; D050981:Erythroid-Specific DNA-Binding Factors; D050982:GATA1 Transcription Factor; C494688:GATA1 protein, human; C409587:GRAP2 protein, human; D011518:Proto-Oncogene Proteins; D050783:Proto-Oncogene Proteins c-ets; D012333:RNA, Messenger; D011993:Recombinant Fusion Proteins; D014157:Transcription Factors; D008156:Luciferases; D001616:beta-Galactosidase; D013755:Tetradecanoylphorbol Acetate