Minimal promoter components of the human growth/differentiation factor-5 gene.
Growth/differentiation factor-5 (GDF-5) is a new member of the BMP family supposed to be involved in chondrogenesis. We cloned the human GDF-5 gene from lambda phage library and sequenced its 3.5-kb 5'-flanking region. The transcription start site was mapped by 5'-RACE to the sequence that coincides with the initiator element. Electrophoresis mobility shift assays (EMSA) demonstrated a zinc finger transcription factor, YY1, to bind to the sequence surrounding the transcription start site. To localize positive and negative regulatory elements in the GDF-5 5'-upstream region, we constructed a series of progressively deleted promoter-reporter plasmids. The transient transfection assay with human osteoblastic Hos cells indicated that a minimal enhancer element resides within -117 to -27 relative to the transcription initiation site. Since the GDF-5 promoter was active even in fibroblastic L cells, a mechanism governing its chondrocyte-restrictive expression needs to be explored.
Sugiura
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Laboratory of Bone Research, Discovery Research Laboratories, Hoechst Marion Roussel Ltd., 3-2, Minamidai 1-chome, Kawagoe, Saitama, 350-1165, Japan. sugiuy79@daiichipharm.co.jp
Hötten
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Kawai
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D020121:5' Untranslated Regions / Q000235:genetics; D001483:Base Sequence; D019485:Bone Morphogenetic Proteins; D002460:Cell Line; D002467:Cell Nucleus / Q000378:metabolism; D003001:Cloning, Molecular; D004268:DNA-Binding Proteins / Q000378:metabolism; D004742:Enhancer Elements, Genetic; D050981:Erythroid-Specific DNA-Binding Factors; D015723:Gene Library; D017930:Genes, Reporter; D055428:Growth Differentiation Factor 5; D006133:Growth Substances / Q000235:genetics*; D006801:Humans; D008969:Molecular Sequence Data; D011401:Promoter Regions, Genetic; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis; D015183:Restriction Mapping; D014157:Transcription Factors / Q000378:metabolism; D014158:Transcription, Genetic; D014162:Transfection; D016212:Transforming Growth Factor beta / Q000235:genetics; D050986:YY1 Transcription Factor; D016335:Zinc Fingers
D020121:5' Untranslated Regions; D019485:Bone Morphogenetic Proteins; D004268:DNA-Binding Proteins; D050981:Erythroid-Specific DNA-Binding Factors; C525893:GDF5 protein, human; D055428:Growth Differentiation Factor 5; D006133:Growth Substances; D011993:Recombinant Fusion Proteins; D014157:Transcription Factors; D016212:Transforming Growth Factor beta; D050986:YY1 Transcription Factor; C494698:YY1 protein, human