1: Cell. 2001 May 4;105(3):403-14. Silenced chromatin is permissive to activator binding and PIC recruitment. Sekinger EA, Gross DS. Department of Biochemistry and Molecular Biology, Louisiana State University Health Sciences Center, Shreveport, LA 71130, USA. Chromatin is thought to repress transcription by limiting access of the DNA to transcription factors. Using a yeast heat shock gene flanked by mating-type silencers as a model system, we find that repressive, SIR-generated heterochromatin is permissive to the constitutive binding of an activator, HSF, and two components of the preinitiation complex (PIC), TBP and Pol II. These factors cohabitate the promoter with Sir silencing proteins and deacetylated nucleosomal histones. The heterochromatic HMRa1 promoter is also occupied by TBP and Pol II, suggesting that SIR regulates gene expression not by restricting factor access to DNA but rather by blocking a step downstream of PIC recruitment. Interestingly, activation of silent promoter chromatin occurs in the absence of histone displacement and without change in histone acetylation state. PMID: 11348596 [PubMed - indexed for MEDLINE] --------------------------------------------------------------- 2: Cell Stress Chaperones. 2000 Jul;5(3):229-42. Potential targets for HSF1 within the preinitiation complex. Yuan CX, Gurley WB. Department of Microbiology and Cell Science, Program in Plant Molecular and Cellular Biology, University of Florida, Gainesville 32611-0700, USA. Protein-protein interactions between human heat shock transcription factor 1 (hHSF1) and general transcription factors TFIIA-gamma, TFIIB, TBP, TAF(II)32, and TAF(II)55 and positive coactivator PC4 were characterized in order to identify potential targets of contact in the transcriptional preinitiation complex. These contacts represent one of the final steps in the signal transfer of heat stress to the transcriptional apparatus. TATA-binding protein (TBP) and transcription factor IIB (TFIIB) were identified as major targets for HSF1 transcriptional activation domains AD1 and AD2 based on in vitro interaction assays. TBP showed affinity for AD2 and a fragment containing AD1, while the core domain of TFIIB interacted primarily with the AD1 fragment. Interactions were also detected between full-length HSF1 and the small subunit (gamma) of TFIIA. PC4 interacted weakly with HSF2 and showed even less affinity for HSF1. Coimmunoprecipitation of transiently expressed TBP in HeLa cells demonstrated that HSF1 AD2 and AD1+AD2 are able to bind TBP in vivo. Assays based on transcriptional interference confirmed predictions that both TBP and TFIIB can interact with HSF1 activation domains in HeLa cells. The negative regulatory region (NR) of HSF1 did not interact with any general factors tested in vitro but did bind TFIID in nuclear extracts through contacts that probably involve TATA associated proteins (TAFs). These results suggest a model for transcriptional regulation by HSF1 that involves a shift between formation of dysfunctional TFIID complexes with the NR and transcriptionally competent complexes with the C-terminal activation domains. PMID: 11005381 [PubMed - indexed for MEDLINE] --------------------------------------------------------------- 3: FEBS Lett. 1998 Oct 9;436(3):318-22. Interaction between the Arabidopsis thaliana heat shock transcription factor HSF1 and the TATA binding protein TBP. Reindl A, Schoffl F. Universitat Tubingen, Biologisches Institut, Lehrstuhl fur Allgemeine Genetik, Germany. The heat shock factor (HSF1) is the central regulator of the heat stress (hs) response and is required for stimulating the transcription of the hs genes and consequently the expression of heat shock proteins. To promote the polymerase II-dependent transcription of the hs genes, HSF has to communicate with the basal transcription machinery. Here, we report that the Arabidopsis thaliana HSF1 interacts directly with TBP, the general TATA box binding transcription factor, as shown by affinity chromatography and electrophoretic mobility shift analyses in vitro. An in vivo interaction between AtHSF1 and AtTBP1 was suggested by results employing the yeast two-hybrid system. PMID: 9801140 [PubMed - indexed for MEDLINE] --------------------------------------------------------------- 4: Mol Cell Biol. 1995 May;15(5):2737-44. Dynamic protein-DNA architecture of a yeast heat shock promoter. Giardina C, Lis JT. Section of Biochemistry, Molecular and Cell Biology, Cornell University, Ithaca, New York 14853, USA. Here we present an in vivo footprinting analysis of the Saccharomyces cerevisiae HSP82 promoter. Consistent with current models, we find that yeast heat shock factor (HSF) binds to strong heat shock elements (HSEs) in non-heat-shocked cells. Upon heat shock, however, additional binding of HSF becomes apparent at weak HSEs of the promoter as well. Recovery from heat shock results in a dramatic reduction in HSF binding at both strong and weak HSEs, consistent with a model in which HSF binding is subject to a negative feedback regulation by heat shock proteins. In vivo KMnO4 footprinting reveals that the interaction of the TATA-binding protein (TBP) with this promoter is also modulated: heat shock slightly increases TBP binding to the promoter and this binding is reduced upon recovery from heat shock. KMnO4 footprinting does not reveal a high density of polymerase at the promoter prior to heat shock, but a large open complex between the transcriptional start site and the TATA box is formed rapidly upon activation, similar to that observed in other yeast genes. PMID: 7739554 [PubMed - indexed for MEDLINE] ---------------------------------------------------------------