Project description:The misrepair of DNA double-strand breaks in close spatial proximity within the nucleus can result in chromosomal rearrangements that are important in the pathogenesis of hematopoietic and solid malignancies. It is unknown why certain epigenetic states, such as those found in stem or progenitor cells, appear to facilitate neoplastic transformation. Here we show that altering the transcriptional state of human astrocytes alters patterns of DNA damage repair from ionizing radiation at a gene locus-specific and genome-wide level. Astrocytes induced into a reactive state exhibited increased DNA repair, compared to non-reactive cells, in actively transcribed chromatin domains after irradiation. In mapping these sites of repair, we identified misrepair events and repair hotspots that were unique to each state. The precise characterization of genomic regions susceptible to mutation in specific epigenetic transcriptional states provides new opportunities for addressing clonal evolution in solid cancers, particularly those where double-strand break induction is a cornerstone of management. Primary astrocyte cells were cultured to confluency and went through a linear monolyer scratch or no scratch.
Project description:The misrepair of DNA double-strand breaks in close spatial proximity within the nucleus can result in chromosomal rearrangements that are important in the pathogenesis of hematopoietic and solid malignancies. It is unknown why certain epigenetic states, such as those found in stem or progenitor cells, appear to facilitate neoplastic transformation. Here we show that altering the transcriptional state of human astrocytes alters patterns of DNA damage repair from ionizing radiation at a gene locus-specific and genome-wide level. Astrocytes induced into a reactive state exhibited increased DNA repair, compared to non-reactive cells, in actively transcribed chromatin domains after irradiation. In mapping these sites of repair, we identified misrepair events and repair hotspots that were unique to each state. The precise characterization of genomic regions susceptible to mutation in specific epigenetic transcriptional states provides new opportunities for addressing clonal evolution in solid cancers, particularly those where double-strand break induction is a cornerstone of management.
Project description:CGH of stage 13 amplifying follicle cells to measure changes in replication fork progression in double-strand break repair mutants Comparative genomic hybridization was performed to compare amplification gradients of stage 13 follicle cells from several double-strand break repair mutants to wild type (OrR) gradients. Two-three replicates were done for each genotype.
2017-02-08 | GSE86012 | GEO
Project description:Repair-seq screens of double-strand break repair
Project description:In the bacterium Escherichia coli, RecG directs DNA synthesis during the repair of DNA double-strand breaks by homologous recombination. Examination of RecA binding during double-strand break repair in Escherichia coli in the presence and absence of RecG protein
Project description:Comparative genomic hybridization was performed to compare amplification gradients of stage 13 follicle cells from several DNA damage checkpoint and double-strand break repair mutants to wild-type (OrR) gradients. Two-three replicates were done for each genotype.
Project description:Counting DNA reads using whole genome sequencing is providing new insight into DNA double-strand break repair (DSBR) in the model organism Escherichia coli. We describe the application of RecA chromatin immunoprecipitation coupled to genomic DNA sequencing (RecA-ChIP-seq) and marker frequency analysis (MFA) to analyse the genomic consequences of DSBR.
Project description:Counting DNA reads using whole genome sequencing is providing new insight into DNA double-strand break repair (DSBR) in the model organism Escherichia coli. We describe the application of RecA chromatin immunoprecipitation coupled to genomic DNA sequencing (RecA-ChIP-seq) and marker frequency analysis (MFA) to analyse the genomic consequences of DSBR.
