title: Cytoplasmic TAF2-TAF8-TAF10 complex provides evidence for nuclear holo-TFIID assembly from preformed submodules.
abstract: 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.
authors.affiliation: 1] European Molecular Biology Laboratory, Grenoble Outstation, 6 rue Jules Horowitz, 38042 Grenoble, France [2] Unit for Virus Host-Cell Interactions, University Grenoble Alpes-EMBL-CNRS, 6 rue Jules Horowitz, 38042 Grenoble, France.
authors.forename: Simon
authors.initials: S
authors.lastname: Trowitzsch
authors.affiliation: 1] European Molecular Biology Laboratory, Grenoble Outstation, 6 rue Jules Horowitz, 38042 Grenoble, France [2] Unit for Virus Host-Cell Interactions, University Grenoble Alpes-EMBL-CNRS, 6 rue Jules Horowitz, 38042 Grenoble, France.
authors.forename: Cristina
authors.initials: C
authors.lastname: Viola
authors.affiliation: Cellular Signaling and Nuclear Dynamics Program, Institut de Génétique et de Biologie Moléculaire et Cellulaire, UMR 7104, INSERM U964, 1 rue Laurent Fries, 67404 Illkirch, France.
authors.forename: Elisabeth
authors.initials: E
authors.lastname: Scheer
authors.affiliation: Cellular Signaling and Nuclear Dynamics Program, Institut de Génétique et de Biologie Moléculaire et Cellulaire, UMR 7104, INSERM U964, 1 rue Laurent Fries, 67404 Illkirch, France.
authors.forename: Sascha
authors.initials: S
authors.lastname: Conic
authors.affiliation: Proteomics Platform, Institut de Génétique et de Biologie Moléculaire et Cellulaire, UMR 7104, INSERM U964, 1 rue Laurent Fries, 67404 Illkirch, France.
authors.forename: Virginie
authors.initials: V
authors.lastname: Chavant
authors.affiliation: Cellular Signaling and Nuclear Dynamics Program, Institut de Génétique et de Biologie Moléculaire et Cellulaire, UMR 7104, INSERM U964, 1 rue Laurent Fries, 67404 Illkirch, France.
authors.forename: Marjorie
authors.initials: M
authors.lastname: Fournier
authors.affiliation: Integrated Structural Biology Department, Institut de Génétique et de Biologie Moléculaire et Cellulaire, UMR 7104, INSERM U964, 1 rue Laurent Fries, 67404 Illkirch, France.
authors.forename: Gabor
authors.initials: G
authors.lastname: Papai
authors.affiliation: Chemistry Research Laboratory, University of Oxford, South Parks Road, Oxford OX1 3TA, UK.
authors.forename: Ima-Obong
authors.initials: IO
authors.lastname: Ebong
authors.affiliation: 1] European Molecular Biology Laboratory, Grenoble Outstation, 6 rue Jules Horowitz, 38042 Grenoble, France [2] Unit for Virus Host-Cell Interactions, University Grenoble Alpes-EMBL-CNRS, 6 rue Jules Horowitz, 38042 Grenoble, France.
authors.forename: Christiane
authors.initials: C
authors.lastname: Schaffitzel
authors.affiliation: Wellcome Trust Centre for Cell Biology, University of Edinburgh, Mayfield Road, Edinburgh EH9 3JR, UK.
authors.forename: Juan
authors.initials: J
authors.lastname: Zou
authors.affiliation: 1] European Molecular Biology Laboratory, Grenoble Outstation, 6 rue Jules Horowitz, 38042 Grenoble, France [2] Unit for Virus Host-Cell Interactions, University Grenoble Alpes-EMBL-CNRS, 6 rue Jules Horowitz, 38042 Grenoble, France.
authors.forename: Matthias
authors.initials: M
authors.lastname: Haffke
authors.affiliation: 1] Wellcome Trust Centre for Cell Biology, University of Edinburgh, Mayfield Road, Edinburgh EH9 3JR, UK [2] Institute of Bioanalytics, Department of Biotechnology, Technische Universität Berlin, 13353 Berlin, Germany.
authors.forename: Juri
authors.initials: J
authors.lastname: Rappsilber
authors.affiliation: Chemistry Research Laboratory, University of Oxford, South Parks Road, Oxford OX1 3TA, UK.
authors.forename: Carol V
authors.initials: CV
authors.lastname: Robinson
authors.affiliation: Integrated Structural Biology Department, Institut de Génétique et de Biologie Moléculaire et Cellulaire, UMR 7104, INSERM U964, 1 rue Laurent Fries, 67404 Illkirch, France.
authors.forename: Patrick
authors.initials: P
authors.lastname: Schultz
authors.affiliation: Cellular Signaling and Nuclear Dynamics Program, Institut de Génétique et de Biologie Moléculaire et Cellulaire, UMR 7104, INSERM U964, 1 rue Laurent Fries, 67404 Illkirch, France.
authors.forename: Laszlo
authors.initials: L
authors.lastname: Tora
authors.affiliation: 1] European Molecular Biology Laboratory, Grenoble Outstation, 6 rue Jules Horowitz, 38042 Grenoble, France [2] Unit for Virus Host-Cell Interactions, University Grenoble Alpes-EMBL-CNRS, 6 rue Jules Horowitz, 38042 Grenoble, France [3] School of Biochemistry, Bristol University, Bristol BS8 1TD, UK.
authors.forename: Imre
authors.initials: I
authors.lastname: Berger
mesh_terms: D020816:Amino Acid Motifs; D002151:Calorimetry; D002467:Cell Nucleus / Q000378:metabolism; D018360:Crystallography, X-Ray; D003593:Cytoplasm / Q000378:metabolism*; D006367:HeLa Cells; D006657:Histones / Q000737:chemistry; D006801:Humans; D013058:Mass Spectrometry / Q000379:methods; D008958:Models, Molecular; D011485:Protein Binding; D055503:Protein Multimerization; D017434:Protein Structure, Tertiary; D011994:Recombinant Proteins / Q000378:metabolism; D020349:Surface Plasmon Resonance; D035381:TATA-Binding Protein Associated Factors / Q000378:metabolism*; D035362:Transcription Factor TFIID / Q000737:chemistry* / Q000378:metabolism; D014157:Transcription Factors / Q000378:metabolism
chemical_list: D006657:Histones; D011994:Recombinant Proteins; C501078:SUPT7L protein, human; C089768:TAF10 protein, human; C486221:TAF2 protein, human; C479459:TAF8 protein, human; D035381:TATA-Binding Protein Associated Factors; D035362:Transcription Factor TFIID; D014157:Transcription Factors