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Acta Biochimica et Biophysica Sinica 2009 41(3):237-245; doi:10.1093/abbs/gmp007
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© The Author 2009. Published by ABBS Editorial Office in association with Oxford University Press on behalf of the Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences.

Isolation and functional analysis of the human glioblastoma-specific promoter region of the human GD3 synthase (hST8Sia I) gene

Hyun-Mi Dae1, Haw-Young Kwon1, Nam-Young Kang1, Na-Ree Song1, Kyoung-Sook Kim1, Cheorl-Ho Kim2, Jai-Heon Lee1 and Young-Choon Lee1,*

1 College of Natural Resources and Life Science, BK21 Center for Silver-Bio Industrialization, Dong-A University, Busan 604-714, South Korea
2 Molecular and Cellular Glycobiology Unit, Department of Biological Sciences, SungKyunKwan University, Kyunggi-Do 440-746, South Korea

* Correspondence address. Tel: +82-51-200-7591; Fax: +82-51-200-6536; E-mail: yclee{at}dau.ac.kr


  Abstract

We identified the promoter region of the human GD3 synthase (hST8Sia I) gene to elucidate the mechanism underlying the regulation of hST8Sia I expression in human glioblastoma cells. The 5'-rapid amplification of cDNA end using mRNA prepared from U-87MG cells revealed the presence of transcription start site of hST8Sia I gene, and the 5'-terminal analysis of its product showed that transcription started from 648 nucleotides upstream of the translational initiation site. Functional analysis of the 5'-flanking region of the hST8Sia I gene by transient expression method revealed that the region from –638 to –498 is important for transcriptional activity of the hST8Sia I gene in U-87MG and T98G cells. This region lacks apparent TATA and CAAT boxes, but contains putative binding sites for transcription factors AREB6 and Elk-1. Site-directed mutagenesis and transient transfection assays demonstrated that both AREB6 and Elk-1 elements in this region were required for the promoter activity in U-87MG and T98G cells. These results indicated that both AREB6 and Elk-1 might play an essential role in the transcriptional activity of hST8Sia I gene essential for GD3 synthesis in human glioblastoma cells.

Keywords    promoter; human GD3 synthase (hST8Sia I) gene; glioblastoma; transcription factor

Received: October 20, 2008; Accepted: November 25, 2008
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