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- Cryo-EM Reveals Promoter DNA Binding and Conformational Flexibility of the General Transcription Fac...
Structure, Volume 17, Issue 11, 11 November 2009, Pages 1442-1452
Hans Elmlund, Vera Baraznenok, Tomas Linder, Zsolt Szilagyi, Reza Rofougaran, Anders Hofer, Hans Hebert, Martin Lindahl and Claes M. Gustafsson
SummaryThe general transcription factor IID (TFIID) is required for initiation of RNA polymerase II-dependent transcription at many eukaryotic promoters. TFIID comprises the TATA-binding protein (TBP) and several conserved TBP-associated factors (TAFs). Recognition of the core promoter by TFIID assists assembly of the preinitiation complex. Using cryo-electron microscopy in combination with methods for ab initio single-particle reconstruction and heterogeneity analysis, we have produced density maps of two conformational states of Schizosaccharomyces pombe TFIID, containing and lacking TBP. We report that TBP-binding is coupled to a massive histone-fold domain rearrangement. Moreover, docking of the TBP-TAF1N-terminus atomic structure to the TFIID map and reconstruction of a TAF-promoter DNA complex helps to account for TAF-dependent regulation of promoter-TBP and promoter-TAF interactions.
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- Human TFIID Binds to Core Promoter DNA in a Reorganized Structural State
Cell, Volume 152, Issue 1, 17 January 2013, Pages 120-131
Michael A. Cianfrocco, George A. Kassavetis, Patricia Grob, Jie Fang, Tamar Juven-Gershon, James T. Kadonaga and Eva Nogales
SummaryA mechanistic description of metazoan transcription is essential for understanding the molecular processes that govern cellular decisions. To provide structural insights into the DNA recognition step of transcription initiation, we used single-particle electron microscopy (EM) to visualize human TFIID with promoter DNA. This analysis revealed that TFIID coexists in two predominant and distinct structural states that differ by a 100 Å translocation of TFIID’s lobe A. The transition between these structural states is modulated by TFIIA, as the presence of TFIIA and promoter DNA facilitates the formation of a rearranged state of TFIID that enables promoter recognition and binding. DNA labeling and footprinting, together with cryo-EM studies, were used to map the locations of TATA, Initiator (Inr), motif ten element (MTE), and downstream core promoter element (DPE) promoter motifs within the TFIID-TFIIA-DNA structure. The existence of two structurally and functionally distinct forms of TFIID suggests that the different conformers may serve as specific targets for the action of regulatory factors.
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- In Vitro Analysis of Huntingtin-Mediated Transcriptional Repression Reveals Multiple Transcription F...
Cell, Volume 123, Issue 7, 29 December 2005, Pages 1241-1253
Weiguo Zhai, Hyunkyung Jeong, Libin Cui, Dimitri Krainc and Robert Tjian
SummaryTranscriptional dysregulation has emerged as a potentially important pathogenic mechanism in Huntington's disease, a neurodegenerative disorder associated with polyglutamine expansion in the huntingtin (htt) protein. Here, we report the development of a biochemically defined in vitro transcription assay that is responsive to mutant htt. We demonstrate that both gene-specific activator protein Sp1 and selective components of the core transcription apparatus, including TFIID and TFIIF, are direct targets inhibited by mutant htt in a polyglutamine-dependent manner. The RAP30 subunit of TFIIF specifically interacts with mutant htt both in vitro and in vivo to interfere with formation of the RAP30-RAP74 native complex. Importantly, overexpression of RAP30 in cultured primary striatal cells protects neurons from mutant htt-induced cellular toxicity and alleviates the transcriptional inhibition of the dopamine D2 receptor gene by mutant htt. Our results suggest a mutant htt-directed repression mechanism involving multiple specific components of the basal transcription apparatus.
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Copyright © 2008 Elsevier Inc. All rights reserved.
Molecular Cell, Volume 29, Issue 1, 81-91, 18 January 2008
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Structural Changes in TAF4b-TFIID Correlate with Promoter Selectivity
Wei-Li Liu1, 2, Robert A. Coleman1, 2, Patricia Grob3, 4, David S. King1, 2, Laurence Florens5, Michael P. Washburn5, Kenneth G. Geles6, Joyce L. Yang1, 2, Vincent Ramey3, 4, Eva Nogales3, 4 and Robert Tjian1, 2, ,
1 Howard Hughes Medical Institute, Li Ka-Shing Center for Biomedical and Health Sciences, University of California, Berkeley, Berkeley, CA 94720, USA
2 Molecular and Cell Biology Department, Li Ka-Shing Center for Biomedical and Health Sciences, University of California, Berkeley, Berkeley, CA 94720, USA
3 Howard Hughes Medical Institute, University of California, Berkeley and Lawrence Berkeley National Laboratory, Berkeley, CA 94720, USA
4 Molecular and Cell Biology Department, University of California, Berkeley and Lawrence Berkeley National Laboratory, Berkeley, CA 94720, USA
5 Stowers Institute for Medical Research, 1000 East 50th Street, Kansas City, MO 64110, USA
6 Wyeth Research, Discovery Oncology, Pearl River, NY 10965, USA
- Proper ovarian development requires the cell type-specific transcription factor TAF4b, a subunit of the core promoter recognition complex TFIID. We present the 35 Å structure of a cell type-specific core promoter recognition complex containing TAF4b and TAF4 (4b/4-IID), which is responsible for directing transcriptional synergy between c-Jun and Sp1 at a TAF4b target promoter. As a first step toward correlating potential structure/function relationships of the prototypic TFIID versus 4b/4-IID, we have compared their 3D structures by electron microscopy and single-particle reconstruction. These studies reveal that TAF4b incorporation into TFIID induces an open conformation at the lobe involved in TFIIA and putative activator interactions. Importantly, this open conformation correlates with differential activator-dependent transcription and promoter recognition by 4b/4-IID. By combining functional and structural analysis, we find that distinct localized structural changes in a megadalton macromolecular assembly can significantly alter its activity and lead to a TAF4b-induced reprogramming of promoter specificity.