title: Isolation and functional analysis of the human glioblastoma-specific promoter region of the human GD3 synthase (hST8Sia I) gene.
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.
authors.affiliation: College of Natural Resources and Life Science, BK21 Center for Silver-Bio Industrialization, Dong-A University, Busan 604-714, South Korea.
authors.forename: Hyun-Mi
authors.initials: HM
authors.lastname: Dae
authors.forename: Haw-Young
authors.initials: HY
authors.lastname: Kwon
authors.forename: Nam-Young
authors.initials: NY
authors.lastname: Kang
authors.forename: Na-Ree
authors.initials: NR
authors.lastname: Song
authors.forename: Kyoung-Sook
authors.initials: KS
authors.lastname: Kim
authors.forename: Cheorl-Ho
authors.initials: CH
authors.lastname: Kim
authors.forename: Jai-Heon
authors.initials: JH
authors.lastname: Lee
authors.forename: Young-Choon
authors.initials: YC
authors.lastname: Lee
mesh_terms: D024506:5' Flanking Region; D001483:Base Sequence; D001665:Binding Sites / Q000235:genetics; D001932:Brain Neoplasms / Q000201:enzymology / Q000235:genetics* / Q000473:pathology; D045744:Cell Line, Tumor; D017931:DNA Primers / Q000737:chemistry; D015971:Gene Expression Regulation, Enzymologic; D017930:Genes, Reporter; D005909:Glioblastoma / Q000235:genetics*; D006801:Humans; D049410:Luciferases, Renilla / Q000378:metabolism; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D010957:Plasmids; D011401:Promoter Regions, Genetic; D012333:RNA, Messenger / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012799:Sialyltransferases / Q000235:genetics* / Q000302:isolation & purification*; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism; D024363:Transcription Initiation Site; D014162:Transfection
chemical_list: D017931:DNA Primers; D012333:RNA, Messenger; D014157:Transcription Factors; D049410:Luciferases, Renilla; D012799:Sialyltransferases; C043662:alpha-N-acetylneuraminate alpha-2,8-sialyltransferase