Live cell monitoring of double strand breaks in S. cerevisiae
| العنوان: | Live cell monitoring of double strand breaks in S. cerevisiae |
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| المؤلفون: | Cheng-Sheng Lee, David P. Waterman, Kevin Li, Allison J. Mazzella, Michael Tsabar, James E. Haber, Felix Zhou, Vinay V. Eapen |
| المصدر: | PLoS Genetics, Vol 15, Iss 3, p e1008001 (2019) PLoS Genetics |
| بيانات النشر: | Public Library of Science (PLoS), 2019. |
| سنة النشر: | 2019 |
| مصطلحات موضوعية: | Cancer Research, RAD52, genetic processes, RAD51, Gene Expression, Yeast and Fungal Models, Cell Cycle Proteins, Biochemistry, Microtubules, 0302 clinical medicine, DNA Breaks, Double-Stranded, Homologous Recombination, Cytoskeleton, Genetics (clinical), 0303 health sciences, Chromosome Biology, Eukaryota, Chromatin, Recombinant Proteins, 3. Good health, Cell biology, Nucleic acids, DNA-Binding Proteins, Meiosis, Experimental Organism Systems, Epigenetics, Cellular Structures and Organelles, biological phenomena, cell phenomena, and immunity, Research Article, Saccharomyces cerevisiae Proteins, lcsh:QH426-470, DNA recombination, DNA repair, Green Fluorescent Proteins, Saccharomyces cerevisiae, Biology, Research and Analysis Methods, Saccharomyces, 03 medical and health sciences, Model Organisms, Genetics, Molecular Biology, Mitosis, Ecology, Evolution, Behavior and Systematics, Adaptor Proteins, Signal Transducing, 030304 developmental biology, Biology and life sciences, fungi, Organisms, Fungi, Proteins, DNA, Cell Biology, biology.organism_classification, Yeast, Rad52 DNA Repair and Recombination Protein, Kinetics, enzymes and coenzymes (carbohydrates), lcsh:Genetics, Animal Studies, health occupations, DNA damage, Rad51 Recombinase, Homologous recombination, 030217 neurology & neurosurgery |
| الوصف: | We have used two different live-cell fluorescent protein markers to monitor the formation and localization of double-strand breaks (DSBs) in budding yeast. Using GFP derivatives of the Rad51 recombination protein or the Ddc2 checkpoint protein, we find that cells with three site-specific DSBs, on different chromosomes, usually display 2 or 3 foci that may coalesce and dissociate. This motion is independent of Rad52 and microtubules. Rad51-GFP, by itself, is unable to repair DSBs by homologous recombination in mitotic cells, but is able to form foci and allow repair when heterozygous with a wild type Rad51 protein. The kinetics of formation and disappearance of a Rad51-GFP focus parallels the completion of site-specific DSB repair. However, Rad51-GFP is proficient during meiosis when homozygous, similar to rad51 “site II” mutants that can bind single-stranded DNA but not complete strand exchange. Rad52-RFP and Rad51-GFP co-localize to the same DSB, but a significant minority of foci have Rad51-GFP without visible Rad52-RFP. We conclude that co-localization of foci in cells with 3 DSBs does not represent formation of a homologous recombination “repair center,” as the same distribution of Ddc2-GFP foci was found in the absence of the Rad52 protein. Author summary Double strand breaks (DSBs) pose the greatest threat to the fidelity of an organism’s genome. While much work has been done on the mechanisms of DSB repair, the arrangement and interaction of multiple DSBs within a single cell remain unclear. Using two live-cell fluorescent DSB markers, we show that cells with 3 site-specific DSBs usually form 2 or 3 foci that can may coalesce into fewer foci but also dissociate. The aggregation and mobility of DSBs into a single focus does not depend on the Rad52 recombination protein that is required for various mechanisms of homologous recombination, suggesting that merging of DSBs does not reflect formation of a homologous recombination repair center. |
| اللغة: | English |
| تدمد: | 1553-7404 1553-7390 |
| الوصول الحر: | https://explore.openaire.eu/search/publication?articleId=doi_dedup___::b660708a46084a15bbcb607e10f0d52aTest http://europepmc.org/articles/PMC6415866?pdf=renderTest |
| حقوق: | OPEN |
| رقم الانضمام: | edsair.doi.dedup.....b660708a46084a15bbcb607e10f0d52a |
| قاعدة البيانات: | OpenAIRE |
| تدمد: | 15537404 15537390 |
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