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Xeroderma Pigmentosum Group A Protein Loads as a Separate Factor onto DNA Lesions
Open accessAbstract
Nucleotide excision repair (NER) is the main DNA repair pathway in mammals for removal of UV-induced lesions. NER involves the concerted action of more than 25 polypeptides in a coordinated fashion. The xeroderma pigmentosum group A protein (XPA) has been suggested to function as a central organizer and damage verifier in NER. How XPA reaches DNA lesions and how the protein is distributed in time and space in living cells are unknown. Here we studied XPA in vivo by using a cell line stably expressing physiological levels of functional XPA fused to green fluorescent protein and by applying quantitative fluorescence microscopy. The majority of XPA moves rapidly through the nucleoplasm with a diffusion rate different from those of other NER factors tested, arguing against a preassembled XPA-containing NER complex. DNA damage induced a transient ( approximately 5-min) immobilization of maximally 30% of XPA. Immobilization depends on XPC, indicating that XPA is not the initial lesion recognition protein in vivo. Moreover, loading of replication protein A on NER lesions was not dependent on XPA. Thus, XPA participates in NER by incorporation of free diffusing molecules in XPC-dependent NER-DNA complexes. This study supports a model for a rapid consecutive assembly of free NER factors, and a relatively slow simultaneous disassembly, after repair.
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“Formaldehyde rather than methanol was employed in the reports of a nucleus-only location for XPA [38], [51], [52].”
From UV-Induced Nuclear Import of XPA Is Mediated by Importin-α4 in An ATR-Dependent Manner (2013) · cites this paper as [52]
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“reports of immunofluorescence microscopic analyses that showed that XPA was fully localized to the nucleus even in the absence of DNA damage [38], [51], [52].”
From UV-Induced Nuclear Import of XPA Is Mediated by Importin-α4 in An ATR-Dependent Manner (2013) · cites this paper as [52]
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Metadata
- Authors
- Rademakers, S., Volker, M., Hoogstraten, D., Nigg, A. L., Moné, M. J., van Zeeland, A. A., Hoeijmakers, J. H. J., Houtsmuller, A. B., & Vermeulen, W.
- Affiliations
- Medical Genetics Center (DE)
- Leiden University Medical Center (NL)
- Erasmus MC (NL)
- University of Amsterdam (NL)
- MGC-Department of Radiation Genetics and Chemical Mutagenesis, Leiden University Medical Center, 2333 AL Leiden
- Corresponding author
- Vermeulen, Wim
- Venue
- Molecular and Cellular Biology
- Publisher
- Informa UK Limited
- Published
- 2003-08-01
- Volume
- 23
- Issue
- 16
- Pages
- 5755-5767
- ISSN
- 0270-7306 (ISSN-L)
- 1098-5549
- PMC
- PMC166334
- OpenAlex
- W2165125584
- Cited by
- 149
- References
- 65
- Citations by year
- 2026: 1 · 2025: 1 · 2024: 1 · 2023: 1 · 2022: 4 · 2021: 1 · 2020: 3 · 2019: 5 · 2018: 3 · 2017: 4 · 2016: 6 · 2015: 6 · 2014: 11 · 2013: 3 · 2012: 6
- FWCI
- 4.00
- Type
- journal article
- Language
- en
- Open access
- green
- europepmc.org
- Locations
- Molecular and Cellular Biology (publishedVersion)
- Europe PMC (PubMed Central) (publishedVersion)
- PubMed Central (publishedVersion)
- PubMed (publishedVersion)
- EUR Research Repository (Erasmus University Rotterdam) (submittedVersion, other-oa)
- RePub (Erasmus University Rotterdam) (submittedVersion, other-oa)
- Funders
- Nederlandse Organisatie voor Wetenschappelijk Onderzoek (901-01-229, 805-33-441, 903-68-370)
- KWF Kankerbestrijding
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › DNA Repair Mechanisms
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Nuclear Structure and Function
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Diffusion and Search Dynamics
- Keywords
- XPA, nucleotide excision repair, XPC, replication protein A, DNA damage recognition, UV-induced DNA lesions
- MeSH terms
- Cell Line, Cell Nucleus, DNA Damage, DNA Repair, DNA-Binding Proteins, Dose-Response Relationship, Radiation, Fibroblasts, Humans, Light, Luminescent Proteins, Microscopy, Fluorescence, Models, Biological, Models, Genetic, Peptides, Time Factors, Transfection, Ultraviolet Rays, Immunoblotting, Protein Structure, Tertiary, DNA, Complementary, Microscopy, Confocal, Green Fluorescent Proteins, Xeroderma Pigmentosum Group A Protein
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