Near-atomic resolution visualization of human transcription promoter opening

Yuan He; Chunli Yan; Jie Fang; Carla Inouye; Robert Tjian; Ivaylo Ivanov; Eva Nogales

doi:10.1038/nature17970

Near-atomic resolution visualization of human transcription promoter opening

Nature. 2016 May 19;533(7603):359-65. doi: 10.1038/nature17970. Epub 2016 May 11.

Authors

Yuan He^{1

2}, Chunli Yan³, Jie Fang⁴, Carla Inouye⁵, Robert Tjian^{4

5

6}, Ivaylo Ivanov³, Eva Nogales^{1

4

6}

Affiliations

¹ Molecular Biophysics and Integrative Bio-Imaging Division, Lawrence Berkeley National Laboratory, Berkeley, California 94720, USA.
² Department of Molecular Biosciences, Northwestern University, Evanston, Illinois 60208, USA.
³ Department of Chemistry, Center for Diagnostics and Therapeutics, Georgia State University, Atlanta, Georgia 30302, USA.
⁴ Howard Hughes Medical Institute, University of California, Berkeley, California 94720, USA.
⁵ Li Ka Shing Center for Biomedical and Health Sciences, University of California, Berkeley, California 94720, USA.
⁶ Department of Molecular and Cell Biology, University of California, Berkeley, California 94720, USA.

Abstract

In eukaryotic transcription initiation, a large multi-subunit pre-initiation complex (PIC) that assembles at the core promoter is required for the opening of the duplex DNA and identification of the start site for transcription by RNA polymerase II. Here we use cryo-electron microscropy (cryo-EM) to determine near-atomic resolution structures of the human PIC in a closed state (engaged with duplex DNA), an open state (engaged with a transcription bubble), and an initially transcribing complex (containing six base pairs of DNA-RNA hybrid). Our studies provide structures for previously uncharacterized components of the PIC, such as TFIIE and TFIIH, and segments of TFIIA, TFIIB and TFIIF. Comparison of the different structures reveals the sequential conformational changes that accompany the transition from each state to the next throughout the transcription initiation process. This analysis illustrates the key role of TFIIB in transcription bubble stabilization and provides strong structural support for a translocase activity of XPB.

Publication types

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

MeSH terms

Cryoelectron Microscopy
DNA / chemistry
DNA / metabolism*
DNA / ultrastructure*
DNA Helicases / chemistry
DNA Helicases / metabolism
DNA Helicases / ultrastructure
DNA-Binding Proteins / chemistry
DNA-Binding Proteins / metabolism
DNA-Binding Proteins / ultrastructure
HeLa Cells
Humans
Models, Molecular
Movement*
Multiprotein Complexes / chemistry
Multiprotein Complexes / metabolism*
Multiprotein Complexes / ultrastructure*
Promoter Regions, Genetic*
Protein Conformation
RNA Polymerase II / chemistry
RNA Polymerase II / metabolism
RNA Polymerase II / ultrastructure
Saccharomyces cerevisiae / chemistry
Saccharomyces cerevisiae / metabolism
Transcription Elongation, Genetic
Transcription Factors, TFII / chemistry
Transcription Factors, TFII / metabolism
Transcription Factors, TFII / ultrastructure
Transcription Initiation, Genetic*

Substances

DNA-Binding Proteins
Multiprotein Complexes
Transcription Factors, TFII
XPBC-ERCC-3 protein
DNA
RNA Polymerase II
DNA Helicases

Abstract

Publication types

MeSH terms

Substances

Grants and funding