Dynamic in vivo interaction of DDB2 E3 ubiquitin ligase with UV-damaged DNA is independent of damage-recognition protein XPC
Abstract
Damage DNA binding protein 2 (DDB2) has a high affinity for UV-damaged DNA and has been implicated in the initial steps of global genome nucleotide excision repair (NER) in mammals. DDB2 binds to CUL4A and forms an E3 ubiquitin ligase. In this study, we have analyzed the properties of DDB2 and CUL4A in vivo. The majority of DDB2 and CUL4A diffuse in the nucleus with a diffusion rate consistent with a high molecular mass complex. Essentially all DDB2 binds to UV-induced DNA damage, where each molecule resides for ∼2 minutes. After the induction of DNA damage, DDB2 is proteolytically degraded with a half-life that is two orders of magnitude larger than its residence time on a DNA lesion. This indicates that binding to damaged DNA is not the primary trigger for DDB2 breakdown. The bulk of DDB2 binds to and dissociates from DNA lesions independently of damage-recognition protein XPC. Moreover, the DDB2-containing E3 ubiquitin ligase is bound to many more damaged sites than XPC, suggesting that there is little physical interaction between the two proteins. We propose a scenario in which DDB2 prepares UV-damaged chromatin for assembly of the NER complex.
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The DDB complex initially recognizes CPD lesions and helps recruit XPC.
“The DDB complex initially recognizes the CPD lesions and helps in recruiting XPC”
From ATR- and ATM-Mediated DNA Damage Response Is Dependent on Excision Repair Assembly during G1 but Not in S Phase of Cell Cycle (2016) · cites this paper as [8]
6-4PP lesions are directly recognized by XPC independent of DDB.
“whereas 6-4PP lesions are directly recognized by XPC independent of DDB [5–8].”
From ATR- and ATM-Mediated DNA Damage Response Is Dependent on Excision Repair Assembly during G1 but Not in S Phase of Cell Cycle (2016) · cites this paper as [8]
The DDB complex first recognizes CPD lesions and recruits XPC, whereas XPC can recognize 6-4PP lesions on its own.
“The DDB complex initially recognizes the CPD lesions and recruits XPC [4, 5, 8, 9], whereas XPC can independently recognize 6-4PP lesions [5].”
From NER initiation factors, DDB2 and XPC, regulate UV radiation response by recruiting ATR and ATM kinases to DNA damage sites (2013) · cites this paper as [9]
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- Authors
- Luijsterburg, M. S., Goedhart, J., Moser, J., Kool, H., Geverts, B., Houtsmuller, A. B., Mullenders, L. H. F., Vermeulen, W., & van Driel, R.
- Affiliations
- University of Amsterdam (NL)
- Leiden University Medical Center (NL)
- Erasmus MC (NL)
- Swammerdam Institute for Life Sciences, BioCentrum Amsterdam, Nuclear Organisation Group, University of Amsterdam, Kruislaan 318, 1098 SM Amsterdam, The Netherlands
- ORCID
- Venue
- Journal of Cell Science
- Publisher
- The Company of Biologists
- Published
- 2007-08-01
- Volume
- 120
- Issue
- 15
- Pages
- 2706-2716
- ISSN
- 0021-9533 (ISSN-L)
- 1477-9137
- OpenAlex
- W2153759298
- Cited by
- 105
- References
- 75
- Citations by year
- 2025: 2 · 2024: 2 · 2022: 3 · 2021: 3 · 2020: 6 · 2019: 6 · 2017: 5 · 2016: 6 · 2015: 7 · 2014: 10 · 2013: 7 · 2012: 7
- FWCI
- 4.09
- Type
- journal article
- Language
- en
- Open access
- closed
- Locations
- Journal of Cell Science (publishedVersion)
- PubMed (publishedVersion)
- UvA-DARE (University of Amsterdam) (publishedVersion)
- EUR Research Repository (Erasmus University Rotterdam) (publishedVersion)
- EUR Research Repository (Erasmus University Rotterdam) (submittedVersion)
- University of Groningen research database (University of Groningen / Centre for Information Technology) (submittedVersion)
- Data Archiving and Networked Services (DANS) (submittedVersion)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › DNA Repair Mechanisms
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Ubiquitin and proteasome pathways
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Nuclear Structure and Function
- Keywords
- DDB2, CUL4A, E3 ubiquitin ligase, UV-damaged DNA, nucleotide excision repair, XPC, DNA damage recognition, protein-DNA interactions, protein degradation, nuclear diffusion, chromatin assembly, DNA repair
- MeSH terms
- Animals, Cell Line, DNA, DNA Damage, DNA Repair, DNA-Binding Proteins, Humans, Ultraviolet Rays, Fluorescence Recovery After Photobleaching, Ubiquitin-Protein Ligases, Cullin Proteins, Mice
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