Regulation of AP-1/DNA complex formation in vitro

M C Frame, N M Wilkie, A J Darling, A Chudleigh, A Pintzas, J C Lang, D A Gillespie

Research output: Contribution to journalArticlepeer-review

Abstract

The level of AP-1 DNA-binding activity exhibited in vitro by unfractionated extracts of Hela nuclei can be stimulated by a low molecular weight fraction from rabbit reticulocyte lysate. Stimulation also requires a heat labile component of the nuclear extract, probably a protein. Stimulated and unstimulated extracts with high and low AP-1 DNA-binding activities contain the same levels of proteins reactive with antisera against Jun and Fos, proteins which are shown to be involved in the AP-1/DNA complexes detected in vitro. The low molecular weight fraction from reticulocyte lysate can be substituted by the reducing agent dithiothreitol (DTT) in the stimulation reaction and conversely oxidised glutathione greatly reduces formation of AP-1/DNA complexes. The binding activities of transcription factors SP-1, NF-1 and CBP are unaffected by DTT or oxidised glutathione. These observations, taken together, suggest that the efficiency with which pre-existing Fos and Jun proteins can bind an AP-1 target sequence in vitro can be controlled by a nuclear activity which is sensitive to oxidation/reduction and that this control mechanism is specific for AP-1.
Original languageEnglish
Pages (from-to)205-9
Number of pages5
JournalOncogene
Volume6
Issue number2
Publication statusPublished - Feb 1991

Keywords

  • Animals
  • DNA
  • DNA-Binding Proteins
  • Dithiothreitol
  • HeLa Cells
  • Humans
  • Proto-Oncogene Proteins
  • Proto-Oncogene Proteins c-fos
  • Proto-Oncogene Proteins c-jun
  • Rabbits
  • Transcription Factors

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