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Mechanism of open complex and dual incision formation by human nucleotide excision repair factors
Open accessAbstract
During nucleotide excision repair in human cells, a damaged DNA strand is cleaved by two endonucleases, XPG on the 3' side of the lesion and ERCC1-XPF on the 5' side. These structure-specific enzymes act at junctions between duplex and single-stranded DNA. ATP-dependent formation of an open DNA structure of approximately 25 nt around the adduct precedes this dual incision. We investigated the mechanism of open complex formation and find that mutations in XPB or XPD, the DNA helicase subunits of the transcription and repair factor TFIIH, can completely prevent opening and dual incision in cell-free extracts. A deficiency in XPC protein also prevents opening. The absence of RPA, XPA or XPG activities leads to an intermediate level of strand separation. In contrast, XPF or ERCC1-defective extracts open normally and generate a 3' incision, but fail to form the 5' incision. This same repair defect was observed in extracts from human xeroderma pigmentosum cells with an alteration in the C-terminal domain of XPB, suggesting that XPB has an additional role in facilitating 5' incision by ERCC1-XPF nuclease. These data support a mechanism in which TFIIH-associated helicase activity and XPC protein catalyze initial formation of the key open intermediate, with full extension to the cleavage sites promoted by the other core nucleotide excision repair factors. Opening is followed by dual incision, with the 3' cleavage made first.
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“hHR23B-XPC is absolutely required for dual incision as well as for open complex formation during GG-NER [243, 261].”
From Molecular Mechanisms of Ultraviolet Radiation‐Induced DNA Damage and Repair (2010) · cites this paper as [243]
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Citations
Cites
Mammalian DNA nucleotide excision repair reconstituted with purified protein components
Aboussekhra, A., Biggerstaff, M., Shivji, M. K. K., Vilpo, J. A., Moncollin, V., Podust, V. N., Protić, M., Hübscher, U., Egly, J.-M., & Wood, R. D. (1995) · Cell · doi:10.1016/0092-8674(95)90289-9 · Cited by 837
Self-citationProliferating cell nuclear antigen is required for DNA excision repair
Shivji, M. K. K., Kenny, M. K., & Wood, R. D. (1992) · Cell · doi:10.1016/0092-8674(92)90416-a · Cited by 803
Self-citationDNA Repair Helicase: a Component of BTF2 (TFIIH) Basic Transcription Factor
Schaeffer, L., Roy, R., Humbert, S., Moncollin, V., Vermeulen, W., Hoeijmakers, J. H. J., Chambon, P., & Egly, J. (1993) · Science · doi:10.1126/science.8465201 · Cited by 794
Self-citationXeroderma Pigmentosum Group F Caused by a Defect in a Structure-Specific DNA Repair Endonuclease
Sijbers, A. M., de Laat, W. L., Ariza, R. R., Biggerstaff, M., Wei, Y.-F., Moggs, J. G., Carter, K. C., Shell, B. K., Evans, E. L., de Jong, M. C., et al. (1996) · Cell · doi:10.1016/s0092-8674(00)80155-5 · Cited by 546
Self-citationXPG endonuclease makes the 3′ incision in human DNA nucleotide excision repair
O’Donovan, A., Davies, A. A., Moggs, J. G., West, S. C., & Wood, R. D. (1994) · Nature · doi:10.1038/371432a0 · Cited by 487
Self-citationWeeda, G., van Ham, R. C. A., Vermeulen, W., Bootsma, D., van der Eb, A. J., & Hoeijmakers, J. H. J. (1990) · Cell · doi:10.1016/0092-8674(90)90122-u · Cited by 432
RPA involvement in the damage-recognition and incision steps of nucleotide excision repair
He, Z., Henricksen, L. A., Wold, M. S., & Ingles, C. J. (1995) · Nature · doi:10.1038/374566a0 · Cited by 389
The ERCC2/DNA repair protein is associated with the class II BTF2/TFIIH transcription factor.
Schaeffer, L., Moncollin, V., Roy, R., Staub, A., Mezzina, M., Sarasin, A. R., Weeda, G., Hoeijmakers, J. H. J., & Egly, J. (1994) · The EMBO Journal · doi:10.1002/j.1460-2075.1994.tb06522.x · Cited by 381
Self-citationReaction Mechanism of Human DNA Repair Excision Nuclease
Mu, D., Hsu, D. S., & Sancar, A. (1996) · Journal of Biological Chemistry · doi:10.1074/jbc.271.14.8285 · Cited by 374
Dual roles of a multiprotein complex from S. cerevisiae in transcription and DNA repair
Feaver, W. J., Svejstrup, J. Q., Bardwell, L., Bardwell, A. J., Buratowski, S., Gulyas, K. D., Donahue, T. F., Friedberg, E. C., & Kornberg, R. D. (1993) · Cell · doi:10.1016/0092-8674(93)90624-y · Cited by 319
Cited by
Sorted by citations
Molecular Mechanisms of Mammalian DNA Repair and the DNA Damage Checkpoints
Sancar, A., Lindsey-Boltz, L. A., Ünsal-Kaçmaz, K., & Linn, S. (2004) · Annual Review of Biochemistry · doi:10.1146/annurev.biochem.73.011303.073723 · Cited by 3,317
Molecular Mechanisms of Ultraviolet Radiation‐Induced DNA Damage and Repair
Rastogi, R. P., Richa, Kumar, A., Tyagi, M. B., & Sinha, R. P. (2010) · Journal of Nucleic Acids · doi:10.4061/2010/592980 · Cited by 1,329
“In GG-NER pathway, lesions produced in transcriptionally silent areas of the genome are recognized by hHR23BXPC protein complex in an energy-independent manner.”
BackgroundInfluentialMolecular mechanism of nucleotide excision repair
de Laat, W. L., Jaspers, N. G. J., & Hoeijmakers, J. H. J. (1999) · Genes & Development · doi:10.1101/gad.13.7.768 · Cited by 1,122
Sugasawa, K., Ng, J. M. Y., Masutani, C., Iwai, S., van der Spek, P. J., Eker, A. P. M., Hanaoka, F., Bootsma, D., & Hoeijmakers, J. H. J. (1998) · Molecular Cell · doi:10.1016/s1097-2765(00)80132-x · Cited by 899
Nucleotide Excision Repair in EukaryotesPreprint
Schärer, O. D. (2013) · Cold Spring Harbor Perspectives in Biology · doi:10.1101/cshperspect.a012609 · Cited by 863
Metadata
- Authors
- Evans, E., Moggs, J. G., Hwang, J. R., Egly, J., & Wood, R. D.
- Affiliations
- Institut de génétique et de biologie moléculaire et cellulaire (FR)
- Corresponding author
- Evans, Elizabeth
- Venue
- The EMBO Journal
- Published
- 1997-11-01
- Volume
- 16
- Issue
- 21
- Pages
- 6559-6573
- ISSN
- 0261-4189 (ISSN-L)
- 1460-2075
- PMC
- PMC1170260
- OpenAlex
- W2044000184
- Cited by
- 500
- References
- 51
- Citations by year
- 2026: 9 · 2025: 6 · 2024: 11 · 2023: 12 · 2022: 4 · 2021: 12 · 2020: 13 · 2019: 12 · 2018: 10 · 2017: 9 · 2016: 16 · 2015: 18 · 2014: 14 · 2013: 11 · 2012: 16
- FWCI
- 6.52
- Type
- journal article
- Language
- en
- Open access
- bronze
- doi.org
- Locations
- The EMBO Journal (publishedVersion)
- Europe PMC (PubMed Central) (publishedVersion)
- PubMed Central (publishedVersion)
- PubMed (publishedVersion)
- Funders
- Israel Cancer Research Fund
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › DNA Repair Mechanisms
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › DNA and Nucleic Acid Chemistry
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Nuclear Structure and Function
- Keywords
- nucleotide excision repair, open complex formation, XPG, TFIIH, XPB helicase, XPC, XPD, XPA, RPA, DNA helicase, DNA strand separation, xeroderma pigmentosum, DNA damage response
- MeSH terms
- DNA Excision Repair Protein ERCC-5, Adenosine Triphosphate, Cells, Cultured, Cisplatin, DNA, DNA Damage, DNA Repair, DNA Helicases, DNA-Binding Proteins, DNA, Single-Stranded, Endonucleases, HeLa Cells, Humans, Models, Genetic, Nuclear Proteins, DNA Ligases, Proteins, Substrate Specificity, Transcription Factors, Transcription, Genetic, Xeroderma Pigmentosum, Point Mutation, Transcription Factors, TFII, Macromolecular Substances, Replication Protein A, Transcription Factor TFIIH, Xeroderma Pigmentosum Group D Protein, Xeroderma Pigmentosum Group A Protein
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