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.