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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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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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