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Title 

Expression of the human CMP-NeuAc:GM3 α2,8-sialyltransferase (GD3 synthase) gene through the NF-κB activation in human melanoma SK-MEL-2 cells

Authors 

N Y KangC H KimK S KimJeong Heon KoJ H LeeY K JeongY C Lee

Publisher 

Elsevier

Issue Date 

2007

Citation 

Biochimica et Biophysica Acta (BBA) - Gene Structure and Expression, vol. 1769, no. 11, pp. 622-630

Keywords 

ganglioside GD3human GD3 synthaseSK-MEL-2transcription factorcyclic AMP responsive element binding proteinganglioside gd3 synthaseimmunoglobulin enhancer binding proteinsialyltransferasebinding siteelectrophoretic mobility

Abstract 

To elucidate the mechanism underlying the regulation of human GD3 synthase gene expression in human melanoma SK-MEL-2 cells, we identified the promoter region of the human GD3 synthase gene. The 5′-rapid amplification of cDNA end (5′-RACE) using mRNA prepared from SK-MEL-2 cells revealed the presence of multiple transcription start sites of human GD3 synthase gene. Promoter analyses of the 5′-flanking region of the human GD3 synthase gene using luciferase gene reporter system showed the strong promoter activity in SK-MEL-2 cells. Deletion study revealed that the region as the core promoter from - 1146 to - 646 (A of the translational start ATG as position + 1) was indispensable for endogenous expression of human GD3 synthase gene. This region lacks apparent TATA and CAAT boxes but contains putative binding sites for transcription factors c-Ets-1, CREB, AP-1 and NF-κB. Electrophoretic mobility shift assays using specific competitors, chromatin immunoprecipitation assay and site-directed mutagenesis demonstrated that only NF-κB element in this region is required for the promoter activity in SK-MEL-2 cells. These results indicate that NF-κB plays an essential role in the transcriptional activity of human GD3 synthase gene essential for GD3 synthesis in SK-MEL-2 cells.

ISSN 

0167-4781

Link 

http://dx.doi.org/10.1016/j.bbaexp.2007.08.001

Appears in Collections

1. Journal Articles > Journal Articles

Registered Date

2019-05-02


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