Purification and characterization of a multicomponent AP-1.junD complex from T cells. Dependence on a separate cellular factor for enhanced DNA binding activity

J Biol Chem. 1994 Dec 30;269(52):32963-71.

Abstract

TAP-1 (T-cell AP-1) is a previously identified DNA-binding activity that is rapidly induced in activated T cells in the absence of protein synthesis. This activity has been purified over 2,000-fold from the T-cell line MLA144. Purified TAP-1 is a multicomponent complex composed of 38-kDa and 43-kDa junD polypeptides in association with a separate factor(s), distinct from fos, that partly dissociate from the complex during affinity purification but is required for full TAP-1 DNA-binding activity. When reconstituted with TAP-1, this partly dissociated component strongly enhances the DNA-binding activity of the TAP-1 complex. UV-cross-linking analysis identifies the dissociable component of the TAP-1 complex as a separate class of low molecular mass (23-29-kDa) DNA-binding polypeptide(s). 23-29-kDa polypeptides have been partially purified from nuclear extracts derived from MLA144 that enhance TAP-1 DNA-binding activity over 100-fold and increase its contacts with flanking DNA sequence. These results define TAP-1 as a distinct AP-1.junD-containing complex in T cells whose DNA-binding activity is regulated by the interaction of distinct and separate cellular factor(s).

MeSH terms

  • Animals
  • Base Sequence
  • Biological Factors / metabolism*
  • DNA / metabolism*
  • Hylobates
  • Molecular Sequence Data
  • Molecular Weight
  • Peptides / metabolism
  • Protein Binding
  • Proto-Oncogene Proteins c-jun / isolation & purification*
  • Proto-Oncogene Proteins c-jun / metabolism
  • T-Lymphocytes / metabolism*
  • Transcription Factor AP-1 / isolation & purification*
  • Transcription Factor AP-1 / metabolism
  • Tumor Cells, Cultured
  • Ultraviolet Rays

Substances

  • Biological Factors
  • Peptides
  • Proto-Oncogene Proteins c-jun
  • Transcription Factor AP-1
  • DNA