Identification of glucocorticoid responsive elements (GREs) at far upstream of rat NPY gene.
The location of three glucocorticoid responsive elements (GREs) in rat neuropeptide Y (NPY) gene was determined by chloramphenicol acetyltransferase (CAT) assay and nucleotide sequencing. We have reported that mRNA content of rat prepro-NPY is increased by 1.7-fold in NG108-15 cells by 1 microM dexamethasone, suggesting the presence of GRE in the gene. To identify the element, the 5'-flanking DNA of 3.3 kilobases (kb) was isolated from rat NPY gene. When chimeric chloramphenicol CAT plasmids containing various deletions of the NPY upstream sequence were transfected into NG108-15 cells, the region between -2.9 and -2.1 kb relative to the cap site was found to potentiate the transcription of CAT gene in the presence of 1 microM dexamethasone. The nucleotide sequencing of this region revealed three GRE consensus sequences at -2.5, -2.2 and -2.1 kb. The results indicate that these elements present in the far upstream region of the NPY gene confer induction by glucocorticoids.
Misaki
N
N
Department of Pharmacology I, Osaka University School of Medicine, Japan.
Higuchi
H
H
Yamagata
K
K
Miki
N
N
D000818:Animals; D001483:Base Sequence; D001665:Binding Sites; D015500:Chloramphenicol O-Acetyltransferase / Q000235:genetics / Q000378:metabolism; D003001:Cloning, Molecular; D004247:DNA / Q000235:genetics / Q000302:isolation & purification; D003907:Dexamethasone / Q000494:pharmacology*; D005809:Genes, Regulator / Q000187:drug effects*; D006822:Hybrid Cells; D051379:Mice; D008969:Molecular Sequence Data; D009478:Neuropeptide Y / Q000096:biosynthesis / Q000235:genetics*; D011498:Protein Precursors / Q000096:biosynthesis; D012333:RNA, Messenger / Q000032:analysis / Q000235:genetics / Q000378:metabolism*; D051381:Rats; D014162:Transfection; D014407:Tumor Cells, Cultured
D009478:Neuropeptide Y; D011498:Protein Precursors; D012333:RNA, Messenger; D003907:Dexamethasone; D004247:DNA; C042084:preproneuropeptide Y; D015500:Chloramphenicol O-Acetyltransferase