Cytoplasmic TAF2–TAF8–TAF10 complex provides evidence for nuclear holo–TFIID assembly from preformed submodules

• Published in: Nature communications 2015-01, Vol.6 (1), p.6011-6011, Article 6011
• Main Authors: Trowitzsch, Simon, Viola, Cristina, Scheer, Elisabeth, Conic, Sascha, Chavant, Virginie, Fournier, Marjorie, Papai, Gabor, Ebong, Ima-Obong, Schaffitzel, Christiane, Zou, Juan, Haffke, Matthias, Rappsilber, Juri, Robinson, Carol V., Schultz, Patrick, Tora, Laszlo, Berger, Imre
• Format: Article
• Language: English
• Subjects: 101/28; 101/58; 13/1; 13/106; 13/51; 13/89; 14/35; 14/63; 631/337/572; 631/45/612/1229; 631/45/612/822; 631/80/389/2029; 82/103; 82/75; 82/80; 82/83; Amino Acid Motifs; Calorimetry; Cell Nucleus - metabolism; Crystallography, X-Ray; Cytoplasm - metabolism; HeLa Cells; Histones - chemistry; Humanities and Social Sciences; Humans; Life Sciences; Mass Spectrometry - methods; Models, Molecular; multidisciplinary; Protein Binding; Protein Multimerization; Protein Structure, Tertiary; Recombinant Proteins - metabolism; Science; Science (multidisciplinary); Surface Plasmon Resonance; TATA-Binding Protein Associated Factors - metabolism; Transcription Factor TFIID - chemistry; Transcription Factor TFIID - metabolism; Transcription Factors - metabolism
• ISSN: 2041-1723; 2041-1723
• DOI: 10.1038/ncomms7011

General transcription factor TFIID is a cornerstone of RNA polymerase II transcription initiation in eukaryotic cells. How human TFIID—a megadalton-sized multiprotein complex composed of the TATA-binding protein (TBP) and 13 TBP-associated factors (TAFs)—assembles into a functional transcription factor is poorly understood. Here we describe a heterotrimeric TFIID subcomplex consisting of the TAF2, TAF8 and TAF10 proteins, which assembles in the cytoplasm. Using native mass spectrometry, we define the interactions between the TAFs and uncover a central role for TAF8 in nucleating the complex. X-ray crystallography reveals a non-canonical arrangement of the TAF8–TAF10 histone fold domains. TAF2 binds to multiple motifs within the TAF8 C-terminal region, and these interactions dictate TAF2 incorporation into a core–TFIID complex that exists in the nucleus. Our results provide evidence for a stepwise assembly pathway of nuclear holo–TFIID, regulated by nuclear import of preformed cytoplasmic submodules. TFIID is an essential transcription factor complex that controls the expression of most protein-coding genes in eukaryotes. Here the authors identify and characterize a complex containing TAF2, TAF8 and TAF10, which assembles in the cytoplasm before integration into the nuclear holo–TFIID complex.