Abstract
We studied the transcriptional regulation of human GM3 synthase (hST3Gal V) during megakaryocytic differentiation of K562 cells induced by PMA. Northern blot and reverse transcription polymerase chain reaction (RT-PCR) indicated that the induction of hST3Gal V by phorbol 12-myristate 13-acetate (PMA) is regulated at transcriptional level. To elucidate the mechanism underlying the regulation of the hST3Gal V gene expression during the differentiation of K562 cells induced by PMA, we characterized the promoter region of the hST3Gal V gene. Functional analysis of the 5(')-flanking region of the hST3Gal V gene by transient expression method showed that the -177 to -83 region, which contains a CREB binding site at -143, functions as the PMA-inducible promoter in K562 cells. In addition, gel shift assay and site-directed mutagenesis indicated that the CREB binding site at -143 is crucial for the PMA-induced expression of the hST3Gal V in K562 cells.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Binding Sites
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Cell Differentiation / genetics*
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Cyclic AMP Response Element-Binding Protein / metabolism
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Cyclic AMP Response Element-Binding Protein / physiology*
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Electrophoretic Mobility Shift Assay
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Gene Deletion
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Gene Expression Regulation, Enzymologic / drug effects
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Gene Expression Regulation, Enzymologic / physiology
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Genes, Reporter / genetics
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Humans
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K562 Cells
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Luciferases / genetics
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Megakaryocytes / enzymology*
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Megakaryocytes / pathology
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Mutagenesis, Site-Directed
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Promoter Regions, Genetic / genetics
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RNA, Messenger / analysis
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RNA, Messenger / biosynthesis
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Sialyltransferases / biosynthesis*
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Sialyltransferases / genetics*
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Sialyltransferases / metabolism
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Tetradecanoylphorbol Acetate / pharmacology
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Transcription, Genetic
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Transfection
Substances
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Cyclic AMP Response Element-Binding Protein
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RNA, Messenger
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Luciferases
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Sialyltransferases
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haematoside synthetase
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Tetradecanoylphorbol Acetate