Structure of human TFIID and mechanism of TBP loading onto promoter DNA

Avinash B Patel; Robert K Louder; Basil J Greber; Sebastian Grünberg; Jie Luo; Jie Fang; Yutong Liu; Jeff Ranish; Steve Hahn; Eva Nogales

doi:10.1126/science.aau8872

Structure of human TFIID and mechanism of TBP loading onto promoter DNA

Science. 2018 Dec 21;362(6421):eaau8872. doi: 10.1126/science.aau8872. Epub 2018 Nov 15.

Authors

Avinash B Patel^{1

2}, Robert K Louder^{1

2}, Basil J Greber^{2

3}, Sebastian Grünberg⁴, Jie Luo⁵, Jie Fang⁶, Yutong Liu⁷, Jeff Ranish⁵, Steve Hahn⁴, Eva Nogales^{8

2

6

9}

Affiliations

¹ Biophysics Graduate Group, University of California, Berkeley, CA 94720, USA.
² Molecular Biophysics and Integrative Bio-Imaging Division, Lawrence Berkeley National Laboratory, Berkeley, CA 94720, USA.
³ California Institute for Quantitative Biology (QB3), University of California, Berkeley, CA 94720, USA.
⁴ Division of Basic Sciences, Fred Hutchinson Cancer Research Center, Seattle, WA 98109, USA.
⁵ Institute for Systems Biology, Seattle, WA 98109, USA.
⁶ Howard Hughes Medical Institute, University of California, Berkeley, CA 94720, USA.
⁷ Department of Chemical and Biomolecular Engineering, University of California, Berkeley, CA 94720, USA.
⁸ Biophysics Graduate Group, University of California, Berkeley, CA 94720, USA. enogales@lbl.gov.
⁹ Department of Molecular and Cell Biology, University of California, Berkeley, CA 94720, USA.

Abstract

The general transcription factor IID (TFIID) is a critical component of the eukaryotic transcription preinitiation complex (PIC) and is responsible for recognizing the core promoter DNA and initiating PIC assembly. We used cryo-electron microscopy, chemical cross-linking mass spectrometry, and biochemical reconstitution to determine the complete molecular architecture of TFIID and define the conformational landscape of TFIID in the process of TATA box-binding protein (TBP) loading onto promoter DNA. Our structural analysis revealed five structural states of TFIID in the presence of TFIIA and promoter DNA, showing that the initial binding of TFIID to the downstream promoter positions the upstream DNA and facilitates scanning of TBP for a TATA box and the subsequent engagement of the promoter. Our findings provide a mechanistic model for the specific loading of TBP by TFIID onto the promoter.

Publication types

Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't

MeSH terms

Cross-Linking Reagents / chemistry
Cryoelectron Microscopy
DNA / chemistry
DNA / metabolism
Humans
Promoter Regions, Genetic*
Protein Binding
Protein Domains
Protein Multimerization
Protein Stability
TATA-Box Binding Protein / chemistry*
Transcription Factor TFIID / chemistry*
Transcription Initiation, Genetic*

Substances

Cross-Linking Reagents
TATA-Box Binding Protein
TBP protein, human
Transcription Factor TFIID
DNA

Abstract

Publication types

MeSH terms

Substances

Grants and funding