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PMID: 2315046 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

Rapid repair kinetics of pyrimidine(6-4)pyrimidone photoproducts in human cells are due to excision rather than conformational change.

Nucleic acids research ·Vol. 18 ·No. 4 ·1990-02-25 ·Pages 963-71

Mitchell DL, Brash DE, Nairn RS

Abstract

UV-induced pyrimidine(6-4)pyrimidone photoproducts in DNA of mammalian cells are apparently repaired much more rapidly than cyclobutane dimers. Since only immunological assays for (6-4) photoproducts have been sensitive enough for repair measurements, it was possible that these apparently rapid repair kinetics reflected a change in physical conformation of antibody-binding sites, resulting in epitope loss rather than excision. To discriminate between these possibilities, we developed a procedure to photochemically convert (6-4) photoproducts to single-strand breaks in UV-irradiated DNA with a background low enough to permit repair measurements. Analysis of a specific DNA sequence indicated that photoinduced alkali-labile sites (PALS) were induced with the same site-specificity as (6-4) photoproducts. Normal human and xeroderma pigmentosum (XP) variant cells rapidly excised (6-4) photoproducts measured as PALS, but little repair was seen in cells from XP complementation group A. These repair kinetics corresponded to those determined in the same samples by radioimmunoassay of (6-4) photoproducts. Thus we conclude that the rapid repair of (6-4) photoproducts observed in UV-irradiated human cells is not the result of a conformational change resulting in epitope loss, but reflects excision of this photoproduct from DNA.

MeSH Terms
Cell Line DNA/isolation & purification,radiation effects DNA Repair Dose-Response Relationship, Radiation Endodeoxyribonucleases Humans Hydrolysis Isomerism Kinetics Multienzyme Complexes N-Glycosyl Hydrolases Nucleic Acid Conformation Plasmids Pyrimidine Dimers Ultraviolet Rays Xeroderma Pigmentosum
Chemicals
Multienzyme Complexes Pyrimidine Dimers pyrimidine-pyrimidone dimer UV endonuclease DNA Endodeoxyribonucleases N-Glycosyl Hydrolases
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Mitchell D L
Department of Carcinogenesis, University of Texas M.D. Anderson Cancer Center, Smithville.
Brash D E
Nairn R S
References (29)
29 references, click to expand
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Article Info
Journal
Nucleic acids research
Abbr.
Nucleic Acids Res
ISSN
0305-1048
DOI
10.1093/nar/18.4.963
Published
1990-02-25
Pages
963-71
Language
English
Region
England
NLM ID
0411011
PMCID
PMC330351
Subset
IM
Grants
NCI NIH HHS · CA24540 · United States
NCI NIH HHS · CA36361 · United States
External Links
PubMed source DOI: 10.1093/nar/18.4.963 PMC Full Text View journal details
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