title: Involvement of CREB in the transcriptional regulation of the human GM3 synthase (hST3Gal V) gene during megakaryocytoid differentiation of human leukemia K562 cells.
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.
authors.affiliation: Faculty of Biotechnology, Dong-A University, Busan 604-714, Republic of Korea.
authors.forename: Hee-Jung
authors.initials: HJ
authors.lastname: Choi
authors.forename: Tae-Wook
authors.initials: TW
authors.lastname: Chung
authors.forename: Nam-Young
authors.initials: NY
authors.lastname: Kang
authors.forename: Kyoung-Sook
authors.initials: KS
authors.lastname: Kim
authors.forename: Young-Choon
authors.initials: YC
authors.lastname: Lee
authors.forename: Cheorl-Ho
authors.initials: CH
authors.lastname: Kim
mesh_terms: D001665:Binding Sites; D002454:Cell Differentiation / Q000235:genetics*; D017362:Cyclic AMP Response Element-Binding Protein / Q000378:metabolism / Q000502:physiology*; D024202:Electrophoretic Mobility Shift Assay; D017353:Gene Deletion; D015971:Gene Expression Regulation, Enzymologic / Q000187:drug effects / Q000502:physiology; D017930:Genes, Reporter / Q000235:genetics; D006801:Humans; D020014:K562 Cells; D008156:Luciferases / Q000235:genetics; D008533:Megakaryocytes / Q000201:enzymology* / Q000473:pathology; D016297:Mutagenesis, Site-Directed; D011401:Promoter Regions, Genetic / Q000235:genetics; D012333:RNA, Messenger / Q000032:analysis / Q000096:biosynthesis; D012799:Sialyltransferases / Q000096:biosynthesis* / Q000235:genetics* / Q000378:metabolism; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D014158:Transcription, Genetic; D014162:Transfection
chemical_list: D017362:Cyclic AMP Response Element-Binding Protein; D012333:RNA, Messenger; D008156:Luciferases; D012799:Sialyltransferases; C021343:haematoside synthetase; D013755:Tetradecanoylphorbol Acetate