Identification of NGF-response element in the rat neuropeptide Y gene and induction of the binding proteins.
Gene expression of the rat neuropeptide Y (NPY) increases by 100 times, as the PC12 cells differentiate into sympathetic neuron-like cells with NGF treatment and this increase is partly due to transcriptional activation of the NPY gene (Sabol and Higuchi, Mol. Endocrinol. 4, 384, 1990). To identify the NGF-response element, a transient expression assay was carried out by using the CAT reporter genes containing various lengths of the 5' upstream region of the NPY gene in the PC12 cells. The 48-base element (-80/-33 upstream of the Cap site) was identified as a NGF-response element (NGFRE). Gel shift assay indicated the existence of at least two DNA-binding proteins to NGFRE. The binding activity of the protein(s) (NDF1) to the upper region (-80/-63) was increased by 3-fold with NGF treatment for 24 h. These findings suggest that these nuclear proteins are involved in the enhanced transcription of the NPY gene by NGF.
Higuchi
H
H
Department of Pharmacology I, Osaka University Medical School, Japan.
Nakano
K
K
Miki
N
N
D000818:Animals; D001483:Base Sequence; D001665:Binding Sites; D002454:Cell Differentiation / Q000187:drug effects; D015500:Chloramphenicol O-Acetyltransferase / Q000235:genetics / Q000378:metabolism; D003001:Cloning, Molecular; D008969:Molecular Sequence Data; D009414:Nerve Growth Factors / Q000494:pharmacology*; D009478:Neuropeptide Y / Q000235:genetics*; D009838:Oligodeoxyribonucleotides / Q000302:isolation & purification; D016716:PC12 Cells; D010957:Plasmids; D011401:Promoter Regions, Genetic; D051381:Rats; D017476:Receptors, Neuropeptide Y / Q000096:biosynthesis*; D011994:Recombinant Proteins / Q000378:metabolism; D015183:Restriction Mapping; D017384:Sequence Deletion; D014158:Transcription, Genetic / Q000187:drug effects
D009414:Nerve Growth Factors; D009478:Neuropeptide Y; D009838:Oligodeoxyribonucleotides; D017476:Receptors, Neuropeptide Y; D011994:Recombinant Proteins; D015500:Chloramphenicol O-Acetyltransferase