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.

Basal transcription factors, FRAP, Gene expression, TBP, TFIID
dx.doi.org/10.1242/jcs.064097, hdl.handle.net/1765/85623
Journal of Cell Science
Department of Pathology

De Graaf, P, Mousson, F, Geverts, B, Scheer, E, Tora, L, Houtsmuller, A.B, & Timmers, H.T.M. (2010). Chromatin interaction of TATA-binding protein is dynamically regulated in human cells. Journal of Cell Science, 123(15), 2663–2671. doi:10.1242/jcs.064097