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Promoter proximal pausing in response to stress in serum deprived mouse fibroblasts.

ABSTRACT: We have employed short-capped RNA sequencing (sc-RNA-seq) in order to identify genes whose expression is regulated by promoter proximal pausing of RNA Polymerase II (RNAPI) in response to stress stimulation. We used serum-deprived mouse Swiss 3T3 fibroblasts, either untreated (control) or treated with anisomycin to induce the p38/MAP kinase pathway.

ORGANISM(S): Mus musculus

PROVIDER: GSE55775 | GEO | 2014/08/12

SECONDARY ACCESSION(S): PRJNA240992

REPOSITORIES: GEO

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Promoter proximal pausing in response to stress in serum deprived mouse fibroblasts.

Project description:We have employed short-capped RNA sequencing (sc-RNA-seq) in order to identify genes whose expression is regulated by promoter proximal pausing of RNA Polymerase II (RNAPI) in response to stress stimulation. We used serum-deprived mouse Swiss 3T3 fibroblasts, either untreated (control) or treated with anisomycin to induce the p38/MAP kinase pathway. Serum starved (72 h 0.2% FCS) mouse 3T3 cells were treated with anisomycin (188.5 nM) for 1 h (in duplicates). Untreated, serum-starved cells were used as a control. We isolated nuclear RNA, performed size fractionation followed by isolation of short-capped RNAs (scRNA). scRNAs were subsequently converted into DNA library and sequenced.

2014-08-12 | E-GEOD-55775 | biostudies-arrayexpress

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Transcriptional response to stress in serum deprived mouse fibroblasts

Project description:We have employed gene expression profiling in order to identify targets of transcriptional response to stress in mouse Swiss 3T3 fibroblasts, where we induced p38/MAP kinase pathway using anisomycin. Serum starved (72 h) mouse 3T3 cells were treated with anisomycin (188.5 nM) for 1, 3, and 6 h (in triplicates). Untreated, serum-starved cells were used as a control. RNA was collected and gene expression profiling using mouse Agilent expression array (4x44k) was performed.

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Project description:We have employed chromatin immunoprecipitation followed by high-throughput sequencing (ChIP-seq) to analyze changes in chromatin architecture as well as the occupancy of two RNA Polymerase II (RNAPII) isoforms, initiation-competent (RNAPIIS5ph) as well as elongation-competent (RNAPIIS2ph) upon stress induction. We used resting mouse Swiss 3T3 fibroblasts, either untreated (control) or treated with anisomycin to induce the p38/MAP kinase pathway. Serum starved (72 h 0.2% FCS) mouse 3T3 cells were treated with anisomycin (188.5 nM) for 1 h (in duplicates). Untreated, serum-starved cells were used as a control. Isolated chromatin was subjected to immunoprecipitation with the following antibodies: a-H3S28ph, a-H3K9ac, a-H3K4me3, a-RNAPIIS5ph and a-RNAPIIS2ph. Resulting DNA was sequenced using Illumina platforms.

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