Genomic sequence of human glyoxalase-I: analysis of promoter activity and its regulation.
Glyoxalase-I is a glutathione-binding protein involved in the detoxification of methylglyoxal, a by-product of glycolysis. Aberrations in the expression of human glyoxalase in cancer and diabetes have been reported. To gain a better understanding of the glyoxalase-I regulation under normal physiological conditions and in disease processes, we have cloned 12kb of genomic sequence, comprising five exons, separated by four introns. A fragment comprising 982bp of 5' flanking region was used in the pSEAP reporter system to identify the minimal promoter and to locate any cis-acting functional elements. This region contained a minimal promoter between -20 and -160bp. Cells transfected with a construct containing the 5' flanking sequence exhibited a 45-fold higher activity over vector transfected cells. A twofold reproducible increase in reporter activity was seen with insulin and ZnCl(2) treatments, indicating a functionally operative insulin response element (IRE) and metal response element (MRE). Knowledge regarding the regulation of glyoxalase-I may provide insights into the importance of this enzyme in human diseases.
Ranganathan
S
S
Department of Pharmacology, Fox Chase Cancer Center, 7701 Burholme Avenue, Philadelphia, PA, USA. s_ranganathan@fccc.edu
Ciaccio
P J
PJ
Walsh
E S
ES
Tew
K D
KD
D000469:Alkaline Phosphatase / Q000187:drug effects / Q000235:genetics / Q000378:metabolism; D001483:Base Sequence; D001665:Binding Sites; D002712:Chlorides / Q000494:pharmacology; D003001:Cloning, Molecular; D004247:DNA / Q000737:chemistry / Q000235:genetics*; D003907:Dexamethasone / Q000494:pharmacology; D005091:Exons; D015971:Gene Expression Regulation, Enzymologic; D005796:Genes / Q000235:genetics; D006801:Humans; D007328:Insulin / Q000494:pharmacology; D007438:Introns; D007791:Lactoylglutathione Lyase / Q000235:genetics*; D008969:Molecular Sequence Data; D011401:Promoter Regions, Genetic / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000187:drug effects / Q000235:genetics / Q000378:metabolism; D017422:Sequence Analysis, DNA; D017384:Sequence Deletion; D014158:Transcription, Genetic; D014407:Tumor Cells, Cultured; D017967:Zinc Compounds / Q000494:pharmacology
D002712:Chlorides; D007328:Insulin; D011993:Recombinant Fusion Proteins; D017967:Zinc Compounds; D003907:Dexamethasone; C016837:zinc chloride; D004247:DNA; D000469:Alkaline Phosphatase; D007791:Lactoylglutathione Lyase