Functional analysis of the 5'-flanking region of FTA for expression of rat GDP-L-fucose:beta-D-galactoside 2-alpha-L-fucosyltransferase.
The tissue-specific and species-specific expression of the ABH antigens is well known among vertebrate species and it is regulated by the alpha(1,2)fucosyltransferase that forms the H antigen, a precursor of the A and B antigens. To investigate the mechanisms governing the tissue-specific and species-specific expression of this alpha(1,2)fucosyltransferase, we characterized the gene structure, including the promoter region, of FTA, a rat orthologous homolog of human FUT1 that encodes the H alpha(1, 2)fucosyltransferase and is responsible for the expression of the ABH antigens on human red blood cells. Northern blot and 5'-RACE analyses suggested that at least two forms of FTA mRNA (2.9 and 2.6 kb), which use alternative transcription start sites, are present in the cancer cell lines RCN-9 (rat colon cancer) and PC12 (rat pheochromocytoma), whereas only the 2.6 kb form was detected in normal colon, stomach and pancreas. Transcriptional activity of the 5'-flanking sequence, which contains three putative Sp1-binding sites, but lacks both TATA and CAAT boxes, was examined. Transient transfection experiments of promoter-reporter gene constructs showed high promoter activity in RCN-9, PC12 and human colon cancer (WiDr) cell lines, weak activity in human vascular endothelial (ECV304) cells and no activity in human erythroleukemia (HEL) cells. The results suggest that the 5'-flanking region of FTA contains a tissue-specific promoter. Deletional analysis of the 5'-flanking sequence revealed regions containing cell-type-specific positive acting element(s) and negative regulatory element(s), which are related to the promoter activity.
Soejima
M
M
Division of Human Genetics, Department of Forensic Medicine, Kurume University School of Medicine, Kurume, Japan.
Koda
Y
Y
Wang
B
B
Kimura
H
H
D020121:5' Untranslated Regions; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D019556:COS Cells; D002522:Chlorocebus aethiops; D004730:Endothelium, Vascular / Q000166:cytology / Q000378:metabolism; D004790:Enzyme Induction; D005647:Fucosyltransferases / Q000096:biosynthesis / Q000235:genetics; D005796:Genes; D006367:HeLa Cells; D006801:Humans; D008969:Molecular Sequence Data; D009363:Neoplasm Proteins / Q000096:biosynthesis / Q000235:genetics; D009928:Organ Specificity; D016716:PC12 Cells; D016133:Polymerase Chain Reaction; D011401:Promoter Regions, Genetic; D051381:Rats; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis; D012045:Regulatory Sequences, Nucleic Acid; D017384:Sequence Deletion; D013045:Species Specificity; D014162:Transfection; D000097631:Galactoside 2-alpha-L-fucosyltransferase
D020121:5' Untranslated Regions; D005647:Fucosyltransferases; D009363:Neoplasm Proteins; D011993:Recombinant Fusion Proteins; D000097631:Galactoside 2-alpha-L-fucosyltransferase