Biochemical characterization of the TATA-binding protein-Gal4 activation domain complex.
It has been suggested that complexes between gene-specific activators and the TATA-binding protein (TBP) play an important role in the expression of many genes. However, few detailed studies of well defined activator-TBP complexes have been reported. An analysis of the biochemical properties of the complex formed by the acidic activation domain (AAD) of the yeast activator Gal4 and TBP is presented here. This is shown to be composed of two AAD and one TBP molecule. DNA binding experiments reveal that TATA-containing DNAs and the Gal4 AAD bind TBP competitively, suggesting that the AAD and TATA boxes recognize overlapping surfaces of TBP. The kinetics of the formation and dissociation of the AAD(2)-TBP complex is also probed. The impact of these findings on models for Gal4-mediated transcriptional activation is considered.
Departments of Internal Medicine and Biochemistry, Ryburn Center for Molecular Cardiology, University of Texas Southwestern Medical Center, Dallas, Texas 75390-8573, USA.
Y
Y
Xie
C
C
Denison
S H
SH
Yang
D A
DA
Fancy
T
T
Kodadek
D001667:Binding, Competitive; D003432:Cross-Linking Reagents; D004247:DNA / Q000235:genetics / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000737:chemistry* / Q000378:metabolism*; D005454:Fluorescence Polarization; D005656:Fungal Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D007700:Kinetics; D046911:Macromolecular Substances; D008957:Models, Genetic; D009154:Mutation; D010446:Peptide Fragments / Q000378:metabolism; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000378:metabolism; D029701:Saccharomyces cerevisiae Proteins; D013329:Structure-Activity Relationship; D016385:TATA Box / Q000235:genetics; D035181:TATA-Box Binding Protein; D014157:Transcription Factors / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D015533:Transcriptional Activation; D015003:Yeasts
D003432:Cross-Linking Reagents; D004268:DNA-Binding Proteins; D005656:Fungal Proteins; C052130:GAL4 protein, S cerevisiae; D046911:Macromolecular Substances; D010446:Peptide Fragments; D011993:Recombinant Fusion Proteins; D029701:Saccharomyces cerevisiae Proteins; D035181:TATA-Box Binding Protein; D014157:Transcription Factors; D004247:DNA