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

John F. Bishop, Mario S. Rinaudo, Joseph K. Ritter, Annie C.Y. Chang, Katherine Conant, Donald R. Gehlert

Research output: Contribution to journalArticle

Abstract

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.

Original languageEnglish (US)
Pages (from-to)125-129
Number of pages5
JournalFEBS Letters
Volume264
Issue number1
DOIs
StatePublished - May 7 1990

Keywords

  • Activator protein-2
  • AtT-20 cell
  • DNA binding protein
  • Proopiomelanocortin
  • Transcription factor

ASJC Scopus subject areas

  • Biophysics
  • Structural Biology
  • Biochemistry
  • Molecular Biology
  • Genetics
  • Cell Biology

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  • Cite this

    Bishop, J. F., Rinaudo, M. S., Ritter, J. K., Chang, A. C. Y., Conant, K., & Gehlert, D. R. (1990). A putative AP-2 binding site in the 5' flanking region of the mouse POMC gene. FEBS Letters, 264(1), 125-129. https://doi.org/10.1016/0014-5793(90)80781-D