Letter | Published:

The cryo-electron microscopy structure of human transcription factor IIH

Nature volume 549, pages 414417 (21 September 2017) | Download Citation

Abstract

Human transcription factor IIH (TFIIH) is part of the general transcriptional machinery required by RNA polymerase II for the initiation of eukaryotic gene transcription1. Composed of ten subunits that add up to a molecular mass of about 500 kDa, TFIIH is also essential for nucleotide excision repair1. The seven-subunit TFIIH core complex formed by XPB, XPD, p62, p52, p44, p34, and p8 is competent for DNA repair2, while the CDK-activating kinase subcomplex, which includes the kinase activity of CDK7 as well as the cyclin H and MAT1 subunits, is additionally required for transcription initiation1,2. Mutations in the TFIIH subunits XPB, XPD, and p8 lead to severe premature ageing and cancer propensity in the genetic diseases xeroderma pigmentosum, Cockayne syndrome, and trichothiodystrophy, highlighting the importance of TFIIH for cellular physiology3. Here we present the cryo-electron microscopy structure of human TFIIH at 4.4 Å resolution. The structure reveals the molecular architecture of the TFIIH core complex, the detailed structures of its constituent XPB and XPD ATPases, and how the core and kinase subcomplexes of TFIIH are connected. Additionally, our structure provides insight into the conformational dynamics of TFIIH and the regulation of its activity.

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Acknowledgements

We thank S. Zheng and D. King for providing peptides and XPB monoclonal antibody, P. Grob for cryo-EM support, and T. Houweling and A. Chintangal for computing support. We thank A. B. Patel and R. K. Louder for discussions and help with data collection, and J. H. D. Cate for providing PyMOL scripts. We acknowledge the use of the LAWRENCIUM computing cluster at Lawrence Berkeley National Laboratory and the resources of the National Energy Research Scientific Computing Center, a Department of Energy Office of Science user facility supported by the Office of Science of the US Department of Energy under contract number DE-AC02-05CH11231. This work was funded through NIGMS grants R01-GM63072 to E.N. and P01-GM063210 to P.D.A. B.J.G. was supported by fellowships from the Swiss National Science Foundation (projects P300PA_160983, P300PA_174355), and T.H.D.N. is a University of California, Berkeley Miller Fellow. E.N. is a Howard Hughes medical investigator.

Author information

Affiliations

  1. California Institute for Quantitative Biology (QB3), University of California, Berkeley, California 94720, USA

    • Basil J. Greber
    • , Thi Hoang Duong Nguyen
    •  & Eva Nogales
  2. Molecular Biophysics and Integrative Bio-Imaging Division, Lawrence Berkeley National Laboratory, Berkeley, California 94720, USA

    • Basil J. Greber
    • , Thi Hoang Duong Nguyen
    • , Pavel V. Afonine
    • , Paul D. Adams
    •  & Eva Nogales
  3. Miller Institute for Basic Research in Science, University of California, Berkeley, California 94720, USA

    • Thi Hoang Duong Nguyen
  4. Howard Hughes Medical Institute, University of California, Berkeley, California 94720, USA

    • Jie Fang
    •  & Eva Nogales
  5. Department of Bioengineering, University of California, Berkeley, California 94720, USA

    • Paul D. Adams
  6. Department of Molecular and Cell Biology, University of California, Berkeley, California 94720, USA

    • Eva Nogales

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Contributions

E.N. directed the study. J.F. performed HeLa cell culture and prepared TFIIH. B.J.G. performed cryo-EM specimen preparation, data collection, data processing, and initial model building. B.J.G. and T.H.D.N. built the final model. B.J.G. and T.H.D.N. performed coordinate refinement, supported by P.V.A. in the laboratory of P.D.A. B.J.G. wrote the initial draft of the manuscript and all authors contributed to the final version.

Competing interests

The authors declare no competing financial interests.

Corresponding author

Correspondence to Eva Nogales.

Reviewer Information Nature thanks S. Hahn, O. Llorca and the other anonymous reviewer(s) for their contribution to the peer review of this work.

Publisher's note: Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

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    Supplementary Data

    This file contains an analysis of CX-MS data for human TFIIH.

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https://doi.org/10.1038/nature23903

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