Architecture of the Human and Yeast General Transcription and DNA Repair Factor TFIIH

Jie Luo; Peter Cimermancic; Shruthi Viswanath; Christopher C Ebmeier; Bong Kim; Marine Dehecq; Vishnu Raman; Charles H Greenberg; Riccardo Pellarin; Andrej Sali; Dylan J Taatjes; Steven Hahn; Jeff Ranish

doi:10.1016/j.molcel.2015.07.016

Architecture of the Human and Yeast General Transcription and DNA Repair Factor TFIIH

Mol Cell. 2015 Sep 3;59(5):794-806. doi: 10.1016/j.molcel.2015.07.016.

Authors

Affiliations

¹ Institute for Systems Biology, 401 Terry Avenue North, Seattle, WA 98109, USA.
² Department of Bioengineering and Therapeutic Sciences, Department of Pharmaceutical Chemistry, California Institute for Quantitative Biomedical Sciences, University of California, San Francisco, San Francisco, CA 94158, USA.
³ Department of Chemistry and Biochemistry, University of Colorado, Boulder, CO 80303, USA.
⁴ Division of Basic Sciences, Fred Hutchinson Cancer Research Center, 1100 Fairview Avenue North, PO Box 19024, Mailstop A1-162, Seattle, WA 98109, USA; Génétique des Interactions Macromoléculaires, Institut Pasteur, CNRS UMR3525, 25-28 rue du docteur Roux, 75015 Paris, France.
⁵ Division of Basic Sciences, Fred Hutchinson Cancer Research Center, 1100 Fairview Avenue North, PO Box 19024, Mailstop A1-162, Seattle, WA 98109, USA.
⁶ Institute for Systems Biology, 401 Terry Avenue North, Seattle, WA 98109, USA. Electronic address: jranish@systemsbiology.org.

Abstract

TFIIH is essential for both RNA polymerase II transcription and DNA repair, and mutations in TFIIH can result in human disease. Here, we determine the molecular architecture of human and yeast TFIIH by an integrative approach using chemical crosslinking/mass spectrometry (CXMS) data, biochemical analyses, and previously published electron microscopy maps. We identified four new conserved "topological regions" that function as hubs for TFIIH assembly and more than 35 conserved topological features within TFIIH, illuminating a network of interactions involved in TFIIH assembly and regulation of its activities. We show that one of these conserved regions, the p62/Tfb1 Anchor region, directly interacts with the DNA helicase subunit XPD/Rad3 in native TFIIH and is required for the integrity and function of TFIIH. We also reveal the structural basis for defects in patients with xeroderma pigmentosum and trichothiodystrophy, with mutations found at the interface between the p62 Anchor region and the XPD subunit.

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

Cross-Linking Reagents
DNA Helicases / chemistry
DNA Helicases / genetics
DNA Helicases / metabolism
DNA Repair
Humans
Mass Spectrometry
Models, Molecular
Mutation
Protein Interaction Domains and Motifs
Protein Subunits
Saccharomyces cerevisiae / genetics
Saccharomyces cerevisiae / metabolism
Saccharomyces cerevisiae Proteins / chemistry*
Saccharomyces cerevisiae Proteins / genetics
Saccharomyces cerevisiae Proteins / metabolism*
Transcription Factor TFIIH / chemistry*
Transcription Factor TFIIH / genetics
Transcription Factor TFIIH / metabolism*
Transcription Factors, TFII / chemistry
Transcription Factors, TFII / genetics
Transcription Factors, TFII / metabolism
Transcription, Genetic
Xeroderma Pigmentosum / genetics
Xeroderma Pigmentosum / metabolism
Xeroderma Pigmentosum Group D Protein / chemistry
Xeroderma Pigmentosum Group D Protein / genetics
Xeroderma Pigmentosum Group D Protein / metabolism

Substances

Cross-Linking Reagents
Protein Subunits
Saccharomyces cerevisiae Proteins
TFB1 protein, S cerevisiae
Transcription Factors, TFII
Transcription Factor TFIIH
Rad3 protein, S cerevisiae
DNA Helicases
Xeroderma Pigmentosum Group D Protein
ERCC2 protein, human

Abstract

Publication types

MeSH terms

Substances

Grants and funding