Isolation of coactivators associated with the TATA-binding protein that mediate transcriptional activation

Cell. 1991 Aug 9;66(3):563-76. doi: 10.1016/0092-8674(81)90019-2.


A key step in the regulation of transcription involves interactions between promoter-selective factors and various components of the transcriptional apparatus. Here we report the requirements for transcriptional activation directed by NTF-1, a developmentally regulated transcription factor in Drosophila. Reconstituted transcription with fractionated Drosophila basal factors reveals that activation by NTF-1 requires factors present in the endogenous TFIID fraction that are distinct from the purified TATA-binding protein (TBP). Glycerol gradient sedimentation and immunoprecipitation analyses indicate that TFIID is a multiprotein complex containing TBP and at least six tightly bound TBP-associated factors (TAFs). Preparations of TBP lacking TAFs after fractionation with denaturants no longer support activation by NTF-1 but retain basal level activity. Addition of immunopurified and renatured TAFs to free TBP restores the ability of NTF-1 to activate transcription without influencing basal transcription. These results suggest that one or more of the TAF polypeptides confer coactivator function.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Base Sequence
  • DNA-Binding Proteins / chemistry
  • DNA-Binding Proteins / isolation & purification
  • DNA-Binding Proteins / physiology*
  • Drosophila Proteins
  • Drosophila melanogaster / genetics*
  • Gene Expression Regulation*
  • Hot Temperature
  • Macromolecular Substances
  • Molecular Sequence Data
  • Multiprotein Complexes
  • Oligonucleotides / chemistry
  • Oligonucleotides / metabolism
  • Protein Binding
  • Recombinant Proteins
  • Regulatory Sequences, Nucleic Acid
  • Transcription Factor TFIID
  • Transcription Factors / chemistry
  • Transcription Factors / metabolism*
  • Transcription Factors / physiology*
  • Transcription, Genetic*
  • Urea / chemistry


  • DNA-Binding Proteins
  • Drosophila Proteins
  • Macromolecular Substances
  • Multiprotein Complexes
  • Oligonucleotides
  • Recombinant Proteins
  • Transcription Factor TFIID
  • Transcription Factors
  • grh protein, Drosophila
  • Urea