PloS one, 2019-03, Vol.14 (3), p.e0213383-e0213383
2019
Volltextzugriff (PDF)
Details
- Autor(en) / Beteiligte
- Titel
- PDIP38/PolDIP2 controls the DNA damage tolerance pathways by increasing the relative usage of translesion DNA synthesis over template switching
- Ist Teil von
- PloS one, 2019-03, Vol.14 (3), p.e0213383-e0213383
- Ort / Verlag
- United States: Public Library of Science
- Erscheinungsjahr
- 2019
- Quelle
- Free E-Journal (出版社公開部分のみ）
- Beschreibungen/Notizen
- Replicative DNA polymerases are frequently stalled at damaged template strands. Stalled replication forks are restored by the DNA damage tolerance (DDT) pathways, error-prone translesion DNA synthesis (TLS) to cope with excessive DNA damage, and error-free template switching (TS) by homologous DNA recombination. PDIP38 (Pol-delta interacting protein of 38 kDa), also called Pol δ-interacting protein 2 (PolDIP2), physically associates with TLS DNA polymerases, polymerase η (Polη), Polλ, and PrimPol, and activates them in vitro. It remains unclear whether PDIP38 promotes TLS in vivo, since no method allows for measuring individual TLS events in mammalian cells. We disrupted the PDIP38 gene, generating PDIP38-/- cells from the chicken DT40 and human TK6 B cell lines. These PDIP38-/- cells did not show a significant sensitivity to either UV or H2O2, a phenotype not seen in any TLS-polymerase-deficient DT40 or TK6 mutants. DT40 provides a unique opportunity of examining individual TLS and TS events by the nucleotide sequence analysis of the immunoglobulin variable (Ig V) gene as the cells continuously diversify Ig V by TLS (non-templated Ig V hypermutation) and TS (Ig gene conversion) during in vitro culture. PDIP38-/- cells showed a shift in Ig V diversification from TLS to TS. We measured the relative usage of TLS and TS in TK6 cells at a chemically synthesized UV damage (CPD) integrated into genomic DNA. The loss of PDIP38 also caused an increase in the relative usage of TS. The number of UV-induced sister chromatid exchanges, TS events associated with crossover, was increased a few times in PDIP38-/- human and chicken cells. Collectively, the loss of PDIP38 consistently causes a shift in DDT from TLS to TS without enhancing cellular sensitivity to DNA damage. We propose that PDIP38 controls the relative usage of TLS and TS increasing usage of TLS without changing the overall capability of DDT.
- Sprache
- Englisch
- Identifikatoren
- ISSN: 1932-6203eISSN: 1932-6203DOI: 10.1371/journal.pone.0213383Titel-ID: cdi_plos_journals_2189138988
- Format
- –
- Schlagworte
- Animals, Avian Proteins - deficiency, Avian Proteins - genetics, Avian Proteins - metabolism, Base sequence, Biochemistry, Molecular Biology, Biology and life sciences, Cancer, Cell culture, Cell cycle, Cell Line, Cell lines, Cells (Biology), Chemical damage, Chemical synthesis, Chickens, Chromosomes, Compounds, Crossovers, Damage tolerance, Deoxyribonucleic acid, DNA, DNA - biosynthesis, DNA - genetics, DNA Damage, DNA polymerase, DNA Polymerase beta - deficiency, DNA Polymerase beta - genetics, DNA Polymerase beta - metabolism, DNA polymerases, DNA Primase - deficiency, DNA Primase - genetics, DNA Primase - metabolism, DNA Repair, DNA Replication, DNA synthesis, DNA-directed DNA polymerase, DNA-Directed DNA Polymerase - deficiency, DNA-Directed DNA Polymerase - genetics, DNA-Directed DNA Polymerase - metabolism, Gene conversion, Gene Knockout Techniques, Genes, Immunoglobulin, Genetic aspects, Genetic research, Genomes, Genomics, Health sciences, Homologous recombination, Homology, Humans, Hydrogen peroxide, Immunoglobulins, Immunological tolerance, In vivo methods and tests, Life Sciences, Lymphocytes B, Mammalian cells, Medical research, Medicine, Multifunctional Enzymes - deficiency, Multifunctional Enzymes - genetics, Multifunctional Enzymes - metabolism, Mutants, Neomycin, Nuclear Proteins - deficiency, Nuclear Proteins - genetics, Nuclear Proteins - metabolism, Nucleotide sequence, Organic chemistry, Pharmacy, Phenotypes, Poultry, Proteins, Replication forks, Research and Analysis Methods, Saccharomyces cerevisiae, Sensitivity, Sensitivity enhancement, Switching, Templates, Genetic, Ultraviolet radiation