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

Nat Commun. 2015 Jan 14;6:6011. doi: 10.1038/ncomms7011.

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Motifs
  • Calorimetry
  • Cell Nucleus / metabolism
  • Crystallography, X-Ray
  • Cytoplasm / metabolism*
  • HeLa Cells
  • Histones / chemistry
  • Humans
  • Mass Spectrometry / methods
  • Models, Molecular
  • Protein Binding
  • Protein Multimerization
  • Protein Structure, Tertiary
  • Recombinant Proteins / metabolism
  • Surface Plasmon Resonance
  • TATA-Binding Protein Associated Factors / metabolism*
  • Transcription Factor TFIID / chemistry*
  • Transcription Factor TFIID / metabolism
  • Transcription Factors / metabolism

Substances

  • Histones
  • Recombinant Proteins
  • SUPT7L protein, human
  • TAF10 protein, human
  • TAF2 protein, human
  • TAF8 protein, human
  • TATA-Binding Protein Associated Factors
  • Transcription Factor TFIID
  • Transcription Factors

Associated data

  • PDB/4WV4
  • PDB/4WV6