Involvement of CREB in the transcriptional regulation of the human GM3 synthase (hST3Gal V) gene during megakaryocytoid differentiation of human leukemia K562 cells.
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.
Choi
Hee-Jung
HJ
Faculty of Biotechnology, Dong-A University, Busan 604-714, Republic of Korea.
Chung
Tae-Wook
TW
Kang
Nam-Young
NY
Kim
Kyoung-Sook
KS
Lee
Young-Choon
YC
Kim
Cheorl-Ho
CH
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
D017362:Cyclic AMP Response Element-Binding Protein; D012333:RNA, Messenger; D008156:Luciferases; D012799:Sialyltransferases; C021343:haematoside synthetase; D013755:Tetradecanoylphorbol Acetate