Cloning and characterization of the proximal promoter region of the mouse glutamate-L-cysteine ligase regulatory subunit gene.
We describe upregulation of the mRNA for the mouse glutamate-cysteine ligase regulatory subunit gene (Glcl-r) in Hepa-1 cells treated with beta-napthoflavone (BNF) and tert-butylhydroquinone (tBHQ). A 2-kb fragment of the proximal promoter region of the gene was cloned and sequenced, and sequence analysis reveals a high degree of homology when compared to the human glutamate-cysteine ligase regulatory subunit gene promoter. Primer extension analysis indicates a major transcription start site 218 bp upstream of the translation start codon in a CpG-rich region, suggesting that transcription is Sp1 mediated. Reporter constructs containing nested deletion fragments of the Glcl-r promoter demonstrate that regulatory elements sufficient for basal and tBHQ-inducible expression lie between -273 and -787 bp relative to the translation start codon and that the distal promoter may contain negative regulatory elements.
Hudson
F N
FN
Department of Environmental Health and Center for Ecogenetics and Environmental Health, University of Washington, 4225 Roosevelt Way NE, Suite 100, Seattle, WA 98105-6099, USA.
Kavanagh
T J
TJ
D000818:Animals; D001483:Base Sequence; D003001:Cloning, Molecular; D004247:DNA / Q000032:analysis; D015971:Gene Expression Regulation, Enzymologic; D005721:Glutamate-Cysteine Ligase / Q000235:genetics*; D006801:Humans; D051379:Mice; D008969:Molecular Sequence Data; D011401:Promoter Regions, Genetic / Q000235:genetics*; D012689:Sequence Homology, Nucleic Acid
D004247:DNA; D005721:Glutamate-Cysteine Ligase