Abstract
Patients with the rare neurodevelopmental repair syndrome known as group A trichothiodystrophy (TTD-A) carry mutations in the gene encoding the p8 subunit of the transcription and DNA repair factor TFIIH. Here we describe the crystal structure of a minimal complex between Tfb5, the yeast ortholog of p8, and the C-terminal domain of Tfb2, the yeast p52 subunit of TFIIH. The structure revealed that these two polypeptides adopt the same fold, forming a compact pseudosymmetric heterodimer via a β-strand addition and coiled coils interactions between terminal α-helices. Furthermore, Tfb5 protects a hydrophobic surface in Tfb2 from solvent, providing a rationale for the influence of p8 in the stabilization of p52 and explaining why mutations that weaken p8–p52 interactions lead to a reduced intracellular TFIIH concentration and a defect in nucleotide-excision repair, a common feature of TTD cells.
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References
Feaver, W.J. et al. Dual roles of a multiprotein complex from S. cerevisiae in transcription and DNA repair. Cell 75, 1379–1387 (1993).
Schaeffer, L. et al. DNA repair helicase: a component of BTF2 (TFIIH) basic transcription factor. Science 260, 58–63 (1993).
Hanawalt, P.C. Subpathways of nucleotide excision repair and their regulation. Oncogene 21, 8949–8956 (2002).
Feaver, W.J. et al. Genes for Tfb2, Tfb3, and Tfb4 subunits of yeast transcription/repair factor IIH. Homology to human cyclin-dependent kinase activating kinase and IIH subunits. J. Biol. Chem. 272, 19319–19327 (1997).
Ranish, J.A. et al. Identification of TFB5, a new component of general transcription and DNA repair factor IIH. Nat. Genet. 36, 707–713 (2004).
Fregoso, M. et al. DNA repair and transcriptional deficiencies caused by mutations in the Drosophila p52 subunit of TFIIH generate developmental defects and chromosome fragility. Mol. Cell. Biol. 27, 3640–3650 (2007).
Kraemer, K.H. et al. Xeroderma pigmentosum, trichothiodystrophy and Cockayne syndrome: a complex genotype-phenotype relationship. Neuroscience 145, 1388–1396 (2007).
Dubaele, S. et al. Basal transcription defect discriminates between xeroderma pigmentosum and trichothiodystrophy in XPD patients. Mol. Cell 11, 1635–1646 (2003).
Coin, F. et al. p8/TTD-A as a repair-specific TFIIH subunit. Mol. Cell 21, 215–226 (2006).
Zhou, Y., Kou, H. & Wang, Z. Tfb5 interacts with Tfb2 and facilitates nucleotide excision repair in yeast. Nucleic Acids Res. 35, 861–871 (2007).
Botta, E. et al. Reduced level of the repair/transcription factor TFIIH in trichothiodystrophy. Hum. Mol. Genet. 11, 2919–2928 (2002).
Giglia-Mari, G. et al. A new, tenth subunit of TFIIH is responsible for the DNA repair syndrome trichothiodystrophy group A. Nat. Genet. 36, 714–719 (2004).
Marinoni, J.C. et al. Cloning and characterization of p52, the fifth subunit of the core of the transcription/DNA repair factor TFIIH. EMBO J. 16, 1093–1102 (1997).
Jawhari, A. et al. p52 mediates XPB function within the transcription/repair factor TFIIH. J. Biol. Chem. 277, 31761–31767 (2002).
Vitorino, M. et al. Solution structure and self-association properties of the p8 TFIIH subunit responsible for trichothiodystrophy. J. Mol. Biol. 368, 473–480 (2007).
Remaut, H. & Waksman, G. Protein-protein interaction through β-strand addition. Trends Biochem. Sci. 31, 436–444 (2006).
Chapados, B.R. et al. A. Structural basis for FEN-1 substrate specificity and PCNA-mediated activation in DNA replication and repair. Cell 116, 39–50 (2004).
Das, D. et al. The HhH domain of the human DNA repair protein XPF forms stable homodimers. Proteins 70, 1551–1563 (2008).
Coin, F., Oksenych, V. & Egly, J.-M. Distinct roles for the XPB/p52 and XPD/p44 subcomplexes of TFIIH in damaged DNA opening during nucleotide excision repair. Mol. Cell 26, 245–256 (2007).
Vermeulen, W. et al. Sublimiting concentration of TFIIH transcription/DNA repair factor causes TTD-A trichothiodystrophy disorder. Nat. Genet. 26, 307–313 (2000).
Balch, W.E., Morimoto, R.I., Dillin, A. & Kelly, J.W. Adapting proteostasis for disease intervention. Science 319, 916–919 (2008).
Giglia-Mari, G. et al. Dynamic interaction of TTDA with TFIIH is stabilized by nucleotide excision repair in living cells. PLoS Biol. 4, e156 (2006).
Otwinowski, Z. & Minor, W. Processing of X-ray diffraction data collected in oscillation mode. Methods Enzymol. 276, 307–326 (1997).
Acknowledgements
We thank F. Coin and M. Zurita for fruitful discussions and D. Moras for strong support and advice. We thank V. Cura, A. Mitschler and R. Sanishvili for help with data collection and staff at the European Synchotron Facility (ESRF, ID14) and Advanced Photon Source (APS, GM/CA CAT, ANL, ID23) for use of synchrotron facilities. We acknowledge G. Zeder-Lutz (Bioacapt, ESBS) for SPR measurements, I. Kolb-Cheynel (IGBMC Baculovirus Facility) for protein production in insect cells and C. Brown for excellent technical assistance. This work was supported by funds from ANR-maladies rares contract ANR-05-MRAR-005-02, the Association de la Recherche sur le Cancer, the Ligue contre le Cancer and the European Commission as SPINE2-complexes contract no. LSHG-CT-2006-031220. D.E.K. was supported by EMBO-LTF.
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D.E.K., A.P. and J.-M.E. designed the research; D.E.K., M.V. and A.P. performed the experiments; D.E.K., J.C. and A.P. solved structures; D.E.K., J.C., J.-M.E. and A.P. wrote the manuscript.
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Supplementary Figures 1–4, Supplementary Tables 1 and 2, and Supplementary Methods (PDF 397 kb)
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Kainov, D., Vitorino, M., Cavarelli, J. et al. Structural basis for group A trichothiodystrophy. Nat Struct Mol Biol 15, 980–984 (2008). https://doi.org/10.1038/nsmb.1478
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DOI: https://doi.org/10.1038/nsmb.1478
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