Architecture of the Human and Yeast General Transcription and DNA Repair Factor TFIIH.
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.
Institute for Systems Biology, 401 Terry Avenue North, Seattle, WA 98109, USA.
Jie
J
Luo
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.
Peter
P
Cimermancic
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.
Shruthi
S
Viswanath
Department of Chemistry and Biochemistry, University of Colorado, Boulder, CO 80303, USA.
Christopher C
CC
Ebmeier
Institute for Systems Biology, 401 Terry Avenue North, Seattle, WA 98109, USA.
Bong
B
Kim
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.
Marine
M
Dehecq
Department of Chemistry and Biochemistry, University of Colorado, Boulder, CO 80303, USA.
Vishnu
V
Raman
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.
Charles H
CH
Greenberg
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.
Riccardo
R
Pellarin
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.
Andrej
A
Sali
Department of Chemistry and Biochemistry, University of Colorado, Boulder, CO 80303, USA.
Dylan J
DJ
Taatjes
Division of Basic Sciences, Fred Hutchinson Cancer Research Center, 1100 Fairview Avenue North, PO Box 19024, Mailstop A1-162, Seattle, WA 98109, USA.
Steven
S
Hahn
Institute for Systems Biology, 401 Terry Avenue North, Seattle, WA 98109, USA. Electronic address: jranish@systemsbiology.org.
Jeff
J
Ranish
D003432:Cross-Linking Reagents; D004265:DNA Helicases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D004260:DNA Repair; D006801:Humans; D013058:Mass Spectrometry; D008958:Models, Molecular; D009154:Mutation; D054730:Protein Interaction Domains and Motifs; D021122:Protein Subunits; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000378:metabolism; D029701:Saccharomyces cerevisiae Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D051758:Transcription Factor TFIIH / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D019473:Transcription Factors, TFII / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D014158:Transcription, Genetic; D014983:Xeroderma Pigmentosum / Q000235:genetics / Q000378:metabolism; D051759:Xeroderma Pigmentosum Group D Protein / Q000737:chemistry / Q000235:genetics / Q000378:metabolism
D003432:Cross-Linking Reagents; D021122:Protein Subunits; D029701:Saccharomyces cerevisiae Proteins; C086993:TFB1 protein, S cerevisiae; D019473:Transcription Factors, TFII; D051758:Transcription Factor TFIIH; C061897:Rad3 protein, S cerevisiae; D004265:DNA Helicases; D051759:Xeroderma Pigmentosum Group D Protein; C497292:ERCC2 protein, human