Chromatin interaction of TATA-binding protein is dynamically regulated in human cells.
Gene transcription in mammalian cells is a dynamic process involving regulated assembly of transcription complexes on chromatin in which the TATA-binding protein (TBP) plays a central role. Here, we investigate the dynamic behaviour of TBP by a combination of fluorescence recovery after photobleaching (FRAP) and biochemical assays using human cell lines of different origin. The majority of nucleoplasmic TBP and other TFIID subunits associate with chromatin in a highly dynamic manner. TBP dynamics are regulated by the joint action of the SNF2-related BTAF1 protein and the NC2 complex. Strikingly, both BTAF1 and NC2 predominantly affect TBP dissociation rates, leaving the association rate unchanged. Chromatin immunoprecipitation shows that BTAF1 negatively regulates TBP and NC2 binding to active promoters. Our results support a model for a BTAF1-mediated release of TBP-NC2 complexes from chromatin.
de Graaf
Petra
P
Department of Physiological Chemistry and Netherlands Proteomic Center, University Medical Centre Utrecht, Universiteitsweg 100, 3584 CG Utrecht, Netherlands.
Mousson
Florence
F
Geverts
Bart
B
Scheer
Elisabeth
E
Tora
Laszlo
L
Houtsmuller
Adriaan B
AB
Timmers
H Th Marc
HT
D002460:Cell Line; D045744:Cell Line, Tumor; D002843:Chromatin / Q000235:genetics / Q000378:metabolism*; D047369:Chromatin Immunoprecipitation; D002850:Chromatography, Gel; D036681:Fluorescence Recovery After Photobleaching; D006367:HeLa Cells; D006801:Humans; D015151:Immunoblotting; D047468:Immunoprecipitation; D035381:TATA-Binding Protein Associated Factors / Q000235:genetics / Q000378:metabolism; D035181:TATA-Box Binding Protein / Q000235:genetics / Q000378:metabolism*; D035362:Transcription Factor TFIID / Q000235:genetics / Q000378:metabolism
C109035:BTAF1 protein, human; D002843:Chromatin; D035381:TATA-Binding Protein Associated Factors; D035181:TATA-Box Binding Protein; D035362:Transcription Factor TFIID