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Detection of the small oligonucleotide products of nucleotide excision repair in UVB-irradiated human skin.

Authors
  • Choi, Jun-Hyuk1
  • Han, Sueji1
  • Kemp, Michael G2
  • 1 Center for Bioanalysis, Korea Research Institute of Standards and Science, Republic of Korea; Department of Bio-Analytical Science, University of Science & Technology, Republic of Korea. , (North Korea)
  • 2 Department of Pharmacology and Toxicology, Wright State University Boonshoft School of Medicine, Dayton, OH 45435, United States. Electronic address: [email protected] , (United States)
Type
Published Article
Journal
DNA repair
Publication Date
Dec 05, 2019
Volume
86
Pages
102766–102766
Identifiers
DOI: 10.1016/j.dnarep.2019.102766
PMID: 31838380
Source
Medline
Keywords
Language
English
License
Unknown

Abstract

UVB radiation results in the formation of potentially mutagenic photoproducts in the DNA of epidermal skin cells. In vitro approaches have demonstrated that the nucleotide excision repair (NER) machinery removes UV photoproducts from DNA in the form of small (∼30-nt-long), excised, damage-containing DNA oligonucleotides (sedDNAs). Though this process presumably takes place in human skin exposed to UVB radiation, sedDNAs have not previously been detected in human skin. Using surgically discarded human skin, we have optimized the detection of the sedDNA products of NER from small amounts of human epidermal tissue ex vivo within minutes of UVB exposure and after UVB doses that normally lead to minimal erythema. Moreover, sedDNA generation was inhibited by treatment of skin explants with spironolactone, which depletes the epidermis of the essential NER protein XPB to mimic the skin of xeroderma pigmentosum patients. Time course experiments revealed that a partially degraded form of the sedDNAs could be readily detected even 12 hours following UVB exposure, which indicates that these repair products are relatively stable in human skin epidermis. Together, these data suggest that sedDNA detection may be a useful assay for determining how genetic, environmental, and other factors influence NER activity in human skin epidermis and whether abnormal sedDNA processing contributes to photosensitive skin disorders. Copyright © 2019 Elsevier B.V. All rights reserved.

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