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Title 

Transcriptional regulation of the human GD3 synthase gene expression in Fas-induced Jurkat T cells: a critical role of transcription factor NF-κB in regulated expression

Authors 

Y KangS K KangY C LeeH J ChoiY S LeeS Y ChoYong Sam KimJeong Heon KoC H Kim

Publisher 

Oxford University Press (OUP)

Issue Date 

2006

Citation 

Glycobiology, vol. 16, no. 5, pp. 375-389

Keywords 

Fas-induced Jukart T cellsGD3 synthaseNF-κBtranscriptional regulationcell extractcomplementary DNAcyclic AMP responsive element binding proteincytosineFas antigenganglioside gd3 synthase

Abstract 

The transcriptional regulation mechanisms involved in the up-regulation of Fas-induced GD3 synthase gene have not yet been elucidated. 5′-Rapid amplification of cDNA end (5′-RACE) using mRNA prepared from Fas-induced Jurkat T cells revealed the presence of multiple transcription start sites of human GD3 synthase gene, and the 5′-end analysis of the longest of its product showed that transcription started from 650 nucleotides upstream of the translational initiation site. Promoter analyses of the 5′-flanking region of the human GD3 synthase gene using luciferase gene reporter system showed strong promoter activity in Fas-induced Jurkat T cells. Deletion study revealed that the region from -1146 to -646 (A of the translational start ATG as position +1) was indispensable for the Fas response. This region lacks apparent TATA and CAAT boxes but contains putative binding sites for transcription factors c-Ets-1, cAMP-responsive element-binding (CREB) protein, activating protein 1 (AP-1), and NF-κB. Base-substitution experiment showed that only the NF-κB-binding site of putative binding sites is required for the maximal expression induced by Fas. Both DNase I footprint and electrophoretic mobility shift assays with the nuclear extract of Fas-induced Jurkat T cells revealed that NF-κB was bound specifically to the probe being mediated by its binding site in the promoter sequence. Taken together, these results indicate that NF-κB plays an essential role in the transcriptional activity of human GD3 synthase gene in Fas-induced Jurkat T cells. In addition, the translocation of NF-κB-binding protein to nucleus by Fas activation is also crucial for the increased expression of the GD3 synthase gene in Fas-activated Jurkat T cells.

ISSN 

0959-6658

Link 

http://dx.doi.org/10.1093/glycob/cwj087

Appears in Collections

1. Journal Articles > Journal Articles

Registered Date

2017-04-19


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