Role of the nucleotide excision repair gene ERCC1 in formation of recombination-dependent rearrangements in mammalian cells
| Autor: | Gerald M. Adair, R. Geoffrey Sargent, Rodney S. Nairn, James L. Meservy, Brian D. Perkins, Zsofia Intody, John H. Wilson, April E. Kilburn |
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| Rok vydání: | 2000 |
| Předmět: |
DNA Repair
DNA repair Adenine Phosphoribosyltransferase Biology Transfection Article Cell Line Substrate Specificity chemistry.chemical_compound Endonuclease Genetics Animals Direct repeat Crossing Over Genetic Repetitive Sequences Nucleic Acid Recombination Genetic Nucleic Acid Heteroduplexes Proteins DNA Endonucleases Molecular biology DNA-Binding Proteins Blotting Southern chemistry biology.protein Nucleic Acid Conformation Homologous recombination Gene Deletion Recombination DNA Damage Heteroduplex Nucleotide excision repair |
| Zdroj: | Nucleic Acids Research. 28:3771-3778 |
| ISSN: | 1362-4962 |
| DOI: | 10.1093/nar/28.19.3771 |
| Popis: | Spontaneous recombination between direct repeats at the adenine phosphoribosyltransferase (APRT) locus in ERCC1-deficient cells generates a high frequency of rearrangements that are dependent on the process of homologous recombination, suggesting that rearrangements are formed by misprocessing of recombination intermediates. Given the specificity of the structure-specific Ercc1/Xpf endonuclease, two potential recombination intermediates are substrates for misprocessing in ERCC1(-) cells: heteroduplex loops and heteroduplex intermediates with non-homologous 3' tails. To investigate the roles of each, we constructed repeats that would yield no heteroduplex loops during spontaneous recombination or that would yield two non-homologous 3' tails after treatment with the rare-cutting endonuclease I-SCE:I. Our results indicate that misprocessing of heteroduplex loops is not the major source of recombination-dependent rearrangements in ERCC1-deficient cells. Our results also suggest that the Ercc1/Xpf endonuclease is required for efficient removal of non-homologous 3' tails, like its Rad1/Rad10 counterpart in yeast. Thus, it is likely that misprocessing of non-homologous 3' tails is the primary source of recombination-dependent rearrangements in mammalian cells. We also find an unexpected effect of ERCC1 deficiency on I-SCE:I-stimulated rearrangements, which are not dependent on homologous recombination, suggesting that the ERCC1 gene product may play a role in generating the rearrangements that arise after I-SCE:I-induced double-strand breaks. |
| Databáze: | OpenAIRE |
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