Post‐Replication Repair of DNA in Ultraviolet‐Irradiated Mammalian Cells: No Gaps in DNA Synthesized Late after Ultraviolet Irradiation
Abstract
Ultraviolet light produces pyrimidine dimers in cellular DNA. In mouse cell lines the great majority of these pyrimidine dimers are not excised, but remain in high molecular weight DNA. Hence the DNA used as template for DNA synthesis in the first generation after ultraviolet irradiation must contain such dimers. In two mouse cell lines (L5178Y and 3T3), alkaline sucrose sedimentation studies showed that the DNA pulse‐labelled shortly after irradiation contained gaps, which by analogy with the situation in Escherichia coli, are presumed to be opposite the pyrimidine dimers. In contrast such gaps could not be detected in DNA pulse‐labelled several hours after irradiation, despite the fact that dimers must have been present on the template strands. A possible explanation is that gaps were still being formed, but were filled in rapidly and were therefore not detected. This hypothesis was tested as follows. Either 1 or 8 h after ultraviolet irradiation cells were pulse‐labelled in the presence of theophylline. This purine inhibits the filling in of gaps presumably by binding to the DNA in the gap. Even in the presence of theophylline, the late‐synthesized DNA was considerably larger than the early‐synthesized DNA. This suggests that at late times after irradiation, gaps were either not formed at all opposite the dimers, or were so transient that theophylline could not bind and inhibit their filling in. A model for synthesis of DNA on templates containing pyrimidine dimers in mammalian cells, based on these findings and other recent results, is discussed.
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Blocked polymerases can leave the lesion and PCNA behind and re-initiate downstream via re-priming, generating numerous ssDNA gaps behind forks that likely cause strong Mec1 activation during S phase in UV-irradiated cells.
“Blocked polymerase can leave the lesion and PCNA behind and re-initiate downstream of the lesion via a re-priming mechanism, generating numerous ssDNA gaps behind the forks, which are likely responsible for the strong activation of Mec1 during S phase in UV-irradiated cells [51,82–85].”
From Mind the gap: keeping UV lesions in check (2011) · cites this paper as [83]
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- Authors
- Lehmann, A. R., & Kirk‐Bell, S.
- Affiliations
- University of Sussex (GB)
- ORCID
- Lehmann, Alan R.
- Venue
- European Journal of Biochemistry
- Publisher
- Wiley
- Published
- 1972-12
- Volume
- 31
- Issue
- 3
- Pages
- 438-445
- ISSN
- 0014-2956 (ISSN-L)
- 1432-1033
- OpenAlex
- W1984436184
- Cited by
- 161
- References
- 25
- Citations by year
- 2026: 1 · 2025: 1 · 2023: 2 · 2022: 2 · 2021: 4 · 2020: 1 · 2019: 1 · 2018: 2 · 2017: 3 · 2016: 1 · 2015: 1 · 2014: 2 · 2013: 1
- FWCI
- 6.65
- Type
- journal article
- Language
- en
- Open access
- closed
- Locations
- European Journal of Biochemistry (publishedVersion)
- PubMed (publishedVersion)
- The Mouseion at the JAXlibrary (Jackson Laboratory) (submittedVersion)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › DNA Repair Mechanisms
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › DNA and Nucleic Acid Chemistry
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Cancer Research › Carcinogens and Genotoxicity Assessment
- Keywords
- postreplication repair, UV irradiation, pyrimidine dimers, DNA replication, gap filling, theophylline, L5178Y cells, 3T3 fibroblasts, DNA synthesis, mammalian cells
- MeSH terms
- Animals, Carbon Isotopes, Cell Line, Centrifugation, Density Gradient, DNA, DNA Repair, DNA Replication, Lymphoma, Models, Biological, Molecular Weight, Pyrimidine Nucleotides, Theophylline, Thymidine, Time Factors, Tritium, Ultraviolet Rays, Mice
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