DNA maintenance following bleomycin-induced strand breaks does not require poly(ADP-ribosyl)ation activation in Drosophila S2 cells |
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| Institution: | 1. Horia Hulubei National Institute of Physics and Nuclear Engineering – IFIN HH, 30 Reactorului St., P.O. Box MG-6, Magurele, Bucharest, Romania;2. Cell Physiology and Pharmacology Department, University of Leicester, Maurice Shock Building, University Road, Leicester LE1 9HN, UK;1. The Keenan Research Centre for Biomedical Science, St Michael''s Hospital, Toronto, ON M5B1W8, Canada;2. Department of Anesthesia, University of Toronto, Toronto, ON, Canada;3. Interdepartmental Division of Critical Care Medicine, University of Toronto, Toronto, ON, Canada;4. State Key Laboratory of Respiratory Disease, Guangzhou Institute of Respiratory Disease, First Affiliated Hospital of Guangzhou Medical University, Guangzhou, China;3. Jiangsu Key Laboratory of Infection and Immunity, Institutes of Biology and Medical Sciences, Soochow University, Suzhou 215006, China;4. Chinese Academy of Sciences Key Laboratory of Molecular Virology and Immunology, Institute Pasteur of Shanghai, Chinese Academy of Sciences, Shanghai 200031, China;1. Institut des Neurosciences Paris-Saclay (Neuro-PSI), UMR 9197, Université Paris Sud, CNRS, Université Paris Saclay, Orsay, 91405, France;2. Department of Medical Genetics, University of Tuebingen, Tuebingen, Germany;3. Centre for Rare Diseases, University of Tuebingen, Tuebingen, Germany;1. Intensive Care Unit, Tours University Hospital, Research Center for Respiratory Diseases, INSERM U1100, University of Tours, Tours, France;2. Epidemiology Unit EpiDcliC, Service of Public Health, Tours University Hospital, Tours, France;4. Teaching Hospital of Nancy, Intensive Care Unit, University of Lorraine, Nancy, France;5. MAVIVH, INSERM U1259, University of Tours, Tours, France |
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| Abstract: | BackgroundPoly-ADP ribosylation (PARylation) is a post translational modification, catalyzed by Poly(ADP-ribose)polymerase (PARP) family. In Drosophila, PARP-I (human PARP-1 ortholog) is considered to be the only enzymatically active isoform. PARylation is involved in various cellular processes such as DNA repair in case of base excision and strand-breaks.ObservationsStrand-breaks (SSB and DSB) are detrimental to cell viability and, in Drosophila, that has a unique PARP family organization, little is known on PARP involvement in the control of strand-breaks repair process. In our study, strands-breaks (SSB and DSB) are chemically induced in S2 Drosophila cells using bleomycin. These breaks are efficiently repaired in S2 cells. During the bleomycin treatment, changes in PARylation levels are only detectable in a few cells, and an increase in PARP-I and PARP-II mRNAs is only observed during the recovery period. These results differ strongly from those obtained with Human cells, where PARylation is strongly activating when DNA breaks are generated. Finally, in PARP knock-down cells, DNA stability is altered but no change in strand-breaks repair can be observed.ConclusionsPARP responses in DNA strands-breaks context are functional in Drosophila model as demonstrated by PARP-I and PARP-II mRNA increases. However, no modification of the global PARylation profile is observed during strand-breaks generation, only changes at cellular levels are detectable. Taking together, these results demonstrate that PARylation process in Drosophila is tightly regulated in the context of strands-breaks repair and that PARP is essential during the maintenance of DNA integrity but dispensable in the DNA repair process. |
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| Keywords: | DNA repair DNA breaks PARP Poly(ADP-ribosyl)ation PARG |
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