Characterization of the interactions of PARP-1 with UV-damaged DNA in vivo and in vitro
Open accessAbstract
The existing methodologies for studying robust responses of poly (ADP-ribose) polymerase-1 (PARP-1) to DNA damage with strand breaks are often not suitable for examining its subtle responses to altered DNA without strand breaks, such as UV-damaged DNA. Here we describe two novel assays with which we characterized the interaction of PARP-1 with UV-damaged DNA in vivo and in vitro. Using an in situ fractionation technique to selectively remove free PARP-1 while retaining the DNA-bound PARP-1, we demonstrate a direct recruitment of the endogenous or exogenous PARP-1 to the UV-lesion site in vivo after local irradiation. In addition, using the model oligonucleotides with single UV lesion surrounded by multiple restriction enzyme sites, we demonstrate in vitro that DDB2 and PARP-1 can simultaneously bind to UV-damaged DNA and that PARP-1 casts a bilateral asymmetric footprint from −12 to +9 nucleotides on either side of the UV-lesion. These techniques will permit characterization of different roles of PARP-1 in the repair of UV-damaged DNA and also allow the study of normal housekeeping roles of PARP-1 with undamaged DNA.
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DDB2 may stimulate catalytic activity of PARP-1 in the presence of UV-damaged DNA.
“Furthermore, DDB2 may stimulate catalytic activity of PARP-1 in the presence of UV-damaged DNA [67].”
From Focus on UV-Induced DNA Damage and Repair—Disease Relevance and Protective Strategies (2020) · cites this paper as [67]
PARP1 is directly recruited to UV-lesion sites following local irradiation.
“PARP1 is directly recruited to UV-lesion sites following local irradiation [67].”
From Focus on UV-Induced DNA Damage and Repair—Disease Relevance and Protective Strategies (2020) · cites this paper as [67]
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“PARP1 is directly recruited to UV-lesion sites following local irradiation [67].”
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- Authors
- Purohit, N. K., Robu, M., Shah, R. G., Geacintov, N. E., & Shah, G. M.
- Affiliations
- Centre hospitalier universitaire de Québec (CA)
- New York University (US)
- Corresponding author
- Shah, Girish M.
- Venue
- Scientific Reports
- Published
- 2016-01-12
- Volume
- 6
- Issue
- 1
- Pages
- 19020-19020
- ISSN
- 2045-2322 (ISSN-L)
- PMC
- PMC4709520
- OpenAlex
- W2236834172
- Cited by
- 26
- References
- 33
- Citations by year
- 2026: 1 · 2025: 2 · 2024: 3 · 2023: 1 · 2022: 3 · 2021: 3 · 2020: 4 · 2019: 4 · 2017: 3 · 2016: 2
- FWCI
- 1.56
- Type
- journal article
- Language
- en
- Open access
- gold
- doi.org
- Locations
- Scientific Reports (publishedVersion, cc-by)
- PubMed Central (publishedVersion, cc-by)
- PubMed (publishedVersion)
- Funders
- Shastri Indo-Canadian Institute
- National Institutes of Health
- Canadian Institutes of Health Research (131569_1)
- Natural Sciences and Engineering Research Council of Canada
- National Cancer Institute (R01CA168469, CA168469)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › DNA Repair Mechanisms
- Health Sciences › Medicine › Oncology › PARP inhibition in cancer therapy
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › DNA and Nucleic Acid Chemistry
- Keywords
- PARP1, UV-damaged DNA, DNA damage response, DDB2, DNA repair
- MeSH terms
- Protein Domains, DNA, DNA Damage, Deoxyribodipyrimidine Photo-Lyase, DNA Repair, DNA-Binding Proteins, Humans, Models, Biological, Poly(ADP-ribose) Polymerases, Chemical Precipitation, Protein Binding, Pyrimidine Dimers, Ultraviolet Rays, Reproducibility of Results, DNA Footprinting, Green Fluorescent Proteins, Biocatalysis
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