A putative AP‐2 binding site in the 5' flanking region of the mouse POMC gene

FEBS Letters

Ritter, Joseph K.³  Rinaudo, Mario S.¹  Chang, Annie C.-Y.²  Gehlert, Donald R.¹  Bishop, John F.¹  Conant, Katherine¹ 

[1] Experimental Therapeutics Branch, NINDS, NIH, Bid. 10/5C103, Bethesda. MD 20892, USA;Department of Genetics, Stanford University Medical Center, Stanford, CA 94305, USA;Human Genetics Branch, NICHD, NIH, Bld. 10/8D43, Bethesda, MD 20892 USA

关键词: Activator protein-2;    Proopiomelanocortin;    AtT-20 cell;    DNA binding protein;    Transcription factor;   

DOI  :  10.1016/0014-5793(90)80781-D

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

Using extracts of AtT-20 cell nuclei, protein binding sites on the POMC gene 5'-flanking region were examined with an exonuclease protection approach. One such binding site, located from −119 to −106 bp upstream from the mouse POMC gene transcription initiation site, which exhibited a close homology to the aetivator protein-2 (AP-2) site [1]. A double-stranded oligonucleotide containing this site was subsequently used in gel shift assays to demonstrate AP-2 consensus sequence binding activity in extracts of AtT-20 cell nuclei. Gel shift competition experiments using both homologous and heterologous competitor DNA sequences revealed that the AP-2 like factor(s) exhibited specific binding to the mouse AP-2 consensus sequence. Furthermore, AP-2 factor binding was also modulated by a CTF/NFl-like factor. Pretreatment of AtT-20 cell nuclear extracts with alkaline phosphatase prior to inclusion in gel shift assays led to a reduction in the intensities of AP-2 factor-specific bands, indicating a potential involvement of protein phosphorylation in AP-2 factor binding in AtT-20 cells.

【 授权许可】

Unknown