Formation of a ternary complex by human XPA, ERCC1, and ERCC4(XPF) excision repair proteins
Open accessAbstract
The xeroderma pigmentosum complementation group A (XP-A) protein, XPA, has recently been expressed in Escherichia coli in a soluble and fully functional form. An affinity column was prepared by linking the XPA protein to a solid support. When HeLa cell-free extract capable of excision repair was applied to the column, > 99.9% of the proteins were in the flow-through. However, the flow-through fraction lacked excision activity. The activity was restored by adding the high salt (1 M KCl) eluate of the column to the flow-through fraction. The XPA protein-bound fraction was tested for specific proteins by an in vitro complementation assay with a panel of cell-free extracts from DNA repair-deficient human and rodent cell lines. The XPA-bound fraction complemented cell-free extracts of excision repair cross-complementing 1 (ERCC-1), ERCC-4 (XP-F), and XP-A mutants. We conclude that the XPA damage recognition protein makes a ternary complex with the ERCC1/ERCC4(XPF) heterodimer with a potential nuclease function.
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“Functionally, XPA is believed to play roles in verifying DNA damage, stabilizing repair intermediates, and recruiting other NER factors to the damaged DNA [1,9,10,11,12,13,14,15,16,17,18].”
BackgroundA new structural insight into XPA–DNA interactions
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- Authors
- Park, C. H., & Sancar, A.
- Affiliations
- University of North Carolina at Chapel Hill (US)
- ORCID
- Sancar, A
- Venue
- Proceedings of the National Academy of Sciences
- Publisher
- National Academy of Sciences
- Published
- 1994-05-24
- Volume
- 91
- Issue
- 11
- Pages
- 5017-5021
- ISSN
- 0027-8424 (ISSN-L)
- 1091-6490
- PMC
- PMC43921
- OpenAlex
- W2091370689
- Cited by
- 189
- References
- 26
- Citations by year
- 2026: 1 · 2025: 2 · 2024: 2 · 2023: 2 · 2022: 4 · 2021: 2 · 2019: 3 · 2018: 6 · 2016: 4 · 2015: 1 · 2014: 4 · 2013: 4 · 2012: 6
- FWCI
- 7.21
- Type
- journal article
- Language
- en
- Open access
- green
- www.ncbi.nlm.nih.gov
- Locations
- Proceedings of the National Academy of Sciences (publishedVersion)
- PubMed Central (publishedVersion)
- PubMed (publishedVersion)
- Carolina Digital Repository (University of North Carolina at Chapel Hill) (submittedVersion)
- UNC Libraries
- Funders
- National Institutes of Health
- National Institute of General Medical Sciences (R01GM032833, GM32833)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › DNA Repair Mechanisms
- Health Sciences › Medicine › Pathology and Forensic Medicine › Genetic factors in colorectal cancer
- Physical Sciences › Materials Science › Materials Chemistry › Enzyme Structure and Function
- Keywords
- XPA, ERCC1, xeroderma pigmentosum, nucleotide excision repair, ternary complex, DNA damage recognition, nuclease activity, complementation assay, cell-free extracts, affinity purification, HeLa cells
- MeSH terms
- Animals, Cell-Free System, Chromatography, Affinity, Cloning, Molecular, DNA Damage, DNA Repair, DNA-Binding Proteins, Endonucleases, Escherichia coli, Exonucleases, Genetic Complementation Test, HeLa Cells, Humans, Protein Binding, Proteins, Xeroderma Pigmentosum Group A Protein
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