Dataset Information

Identification of DNA-damage sensing factors in D.melanogaster

ABSTRACT: In order to identify protein involved in sensing DNA double strand breaks and recruiting the repair machinry, we performed in vitro assembly of chromatin on small circular and linear DNA templates, respectively. Proteins involved in chromatin assembly and sensing of free DNA ends were isolated and subjected to proteomics analysis.

OTHER RELATED OMICS DATASETS IN: GSE126415

INSTRUMENT(S): TripleTOF 5600

ORGANISM(S): Drosophila Melanogaster (fruit Fly)

TISSUE(S): Embryo, Early Embryonic Cell

SUBMITTER: Andreas Schmidt

LAB HEAD: Axel Imhof

PROVIDER: PXD012990 | Pride | 2019-10-14

REPOSITORIES: Pride

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Publications

A Drosophila cell-free system that senses DNA breaks and triggers phosphorylation signalling.

Harpprecht Lisa L Baldi Sandro S Schauer Tamas T Schmidt Andreas A Bange Tanja T Robles Maria S MS Kremmer Elisabeth E Imhof Axel A Becker Peter B PB

Nucleic acids research 20190801 14

Preblastoderm Drosophila embryo development is characterized by fast cycles of nuclear divisions. Extracts from these embryos can be used to reconstitute complex chromatin with high efficiency. We now discovered that this chromatin assembly system contains activities that recognize unprotected DNA ends and signal DNA damage through phosphorylation. DNA ends are initially bound by Ku and MRN complexes. Within minutes, the phosphorylation of H2A.V (homologous to γH2A.X) initiates from DNA breaks a ...[more]

PMID: 31147711

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Project description:The present study aims to explore the role of Rim15 in both physiology and genome wide expression in S. cerevisiae under severe caloric restriction. Non-growing but metabolically active cultures of S. cerevisiae are of major interest for application in industry and as model systems for aging in higher eukaryotes. Using retentostat cultivations, almost non-growing but metabolic active cultures can be obtained resulting from the severe caloric restriction, yet not starvation, yeast experiences. Rim15 plays an important role in several nutrient sensing pathways and is involved in activating stress response and glycogen accumulation upon nutrient shortage. To investigate the role of Rim15 in the extreme robustness and glycogen accumulation of anaerobic retentostat cultures, a rim15 deletion strain is compared with its parental strain under anaerobic calorie restriction on both physiology and transcriptome. Rim15 is described as essential for G0 entry and glycogen accumulation in yeast during diauxic shift and stationary phase. Anaerobic retentostat cultures display many stationary phase characteristics, including increased expression levels of Rim15 target genes, suggesting an important role for Rim15 under these conditions. Comparing a rim15 deletion strain and its parental strain revealed both on transcriptome level and physiology indeed a major role in the acquired robustness, glycogen accumulation, but also maintenance of viability and cell cycle arrest. The severe caloric restriction, but not starving, conditions applied together with a thorough physiological and transcriptome analysis of the cultures shows that Rim15 is essential in fine-tuning cell cycle progression with glucose availability under extreme growth-limiting conditions. To investigate the impact of rim15 deletion under severe calorie restricted conditions, transcriptome of a S. cerevisiae rim15 deletion mutant was monitored during anaerobic retentostat cultivation. Five time points were used, one in the starting point of the retentostat (T0 = chemostat) and four during the course of the retentostat (time points 2, 9, 16 and 20 days). Culture triplicates were performed for T0 in chemostat, while two independent cultures were run for retentostat.

Dataset Information

Identification of DNA-damage sensing factors in D.melanogaster

Publications

A Drosophila cell-free system that senses DNA breaks and triggers phosphorylation signalling.

Similar Datasets

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