The human glutaredoxin gene: determination of its organization, transcription start point, and promoter analysis.
A genomic clone for the human glutaredoxin gene was isolated and sequenced. An intron was located within the coding region and began 211 nt downstream of the initiator codon. Except for this intron, the genomic sequence shares 100% identity to the published glutaredoxin cDNA sequence. A second intron was located in the 3' UTR 6 bp downstream of the terminator codon. The tsp of the glutaredoxin gene was determined by primer extension and confirmed by S1 mapping analysis. Analysis of the 5'-flanking region of the gene revealed that the promoter sequences TATA and CCAAT were 30 and 160 bp upstream, respectively, from the tsp. Other potential transcription factor binding sites included NF-E1, HNF-5, P2II and AP-1. Glutaredoxin promoter constructs inserted into a reporter plasmid for firefly luciferase were transfected into fibroblasts, and luciferase activity was 8-10-fold higher compared with controls lacking glutaredoxin promoter. These data indicate that the promoter region of the isolated glutaredoxin gene is functional.
Park
J B
JB
Molecular and Clinical Nutrition Section, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Bethesda, MD 20892-1372, USA.
Levine
M
M
D000595:Amino Acid Sequence; D001483:Base Sequence; D001665:Binding Sites; D002478:Cells, Cultured; D003001:Cloning, Molecular; D005347:Fibroblasts; D005796:Genes / Q000235:genetics*; D054477:Glutaredoxins; D006801:Humans; D007438:Introns / Q000235:genetics; D008969:Molecular Sequence Data; D010088:Oxidoreductases; D011401:Promoter Regions, Genetic / Q000235:genetics*; D011506:Proteins / Q000235:genetics*; D011993:Recombinant Fusion Proteins; D017422:Sequence Analysis, DNA; D012689:Sequence Homology, Nucleic Acid; D014157:Transcription Factors / Q000378:metabolism; D014158:Transcription, Genetic / Q000235:genetics*
D054477:Glutaredoxins; D011506:Proteins; D011993:Recombinant Fusion Proteins; D014157:Transcription Factors; D010088:Oxidoreductases