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Unknown,Transcriptomics,Genomics,Proteomics

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Functional coordination and HuR-mediated regulation of mRNA stability during T cell activation

ABSTRACT: We evaluated changes in mRNA stability and transcription using 4sU metabolic pulse labeling across a four hour time course following activation of Jurkat T cells with PMA and PHA Measurement of total mRNA (T) and 4sU labeled mRNA (IP) in three biological replicates at five time points: prior to activation (U) and the first four hours after activation (1-4)

ORGANISM(S): Homo sapiens

SUBMITTER: Jeff Blackinton

PROVIDER: E-GEOD-71382 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Functional coordination and HuR-mediated regulation of mRNA stability during T cell activation.

Blackinton Jeff G JG Keene Jack D JD

Nucleic acids research 20151020 1

Global mRNA abundance depends on the balance of synthesis and decay of a population of mRNAs. To account for this balance during activation of T cells, we used metabolic labeling to quantify the contributions of RNA transcription and decay over a 4 h time course during activation of leukemia-derived Jurkat T cells. While prior studies suggested more than half of the changes in mRNA abundance were due to RNA stability, we found a smaller but more interesting population of mRNAs changed stability. ...[more]

PMID: 26490963

Similar Datasets

Functional coordination and HuR-mediated regulation of mRNA stability during T cell activation

Project description:We evaluated changes in mRNA stability and transcription using 4sU metabolic pulse labeling across a four hour time course following activation of Jurkat T cells with PMA and PHA

2015-10-28 | GSE71382 | GEO

Identification of phasin PhaF interaction partners and proteomic analysis of polyhydroxyalkanoate granules in Pseudomonas putida

Project description:Phasin PhaF is a multifunctional protein associated with the surface of polyhydroxyalkanoate (PHA) granules (carbonosomes) that contributes significantly to PHA biogenesis in pseudomonads. PhaF participates in PHA granule stabilization and segregation and its deletion has a notable impact on overall transcriptome, PHA accumulation and cell physiology, suggesting a more extensive function than simply being a simply granule structural protein. We followed a systematic approach to detect potential interactions of PhaF with other components of the cell, which could pinpoint unexplored functions of PhaF in the regulation of PHA production. We determined the PhaF interactome in Pseudomonas putida KT2440 using pull-down-mass spectrometry (PD-MS), in which proteins interacting with PhaF were determined. We separately determined the PHA granule proteome (all proteins associated with PHA granules) in Pseudomonas putida KT2440 using two granule preparation methods.

2021-09-09 | PXD018787 | Pride

Viral nucleases reveal an mRNA degradation-transcription feedback loop in mammalian cells

Project description:Gamma-herpesviruses encode a cytoplasmic mRNA-targeting endonuclease, termed SOX, that cleaves the majority of mRNAs within a cell. Cleaved fragments are subsequently degraded by the cellular mRNA degradation machinery. Here, we reveal that mammalian cells respond to this widespread cytoplasmic mRNA decay by altering levels of RNA polymerase II (RNAPII) transcription in the nucleus. Measurements of both RNAPII recruitment to promoters and nascent mRNA synthesis revealed that the majority of affected genes are transcriptionally repressed in SOX-expressing cells. The transcriptional feedback does not occur in response to the initial endonuclease-induced cleavage, but instead to degradation of the cleaved fragments by cellular exonucleases. In particular, Xrn1 catalytic activity is required for transcriptional repression. Notably, viral mRNA transcription escapes decay-induced repression, and this escape requires Xrn1. Collectively, these results indicate that mRNA decay rates impact transcription in mammalian cells, and that gamma-herpesviruses have incorporated this feedback mechanism into their own gene expression strategy. NIH 3T3 cells were mock, WT, or ∆HS infected with MHV68 in duplicate and 4sU-labeled RNA isolated. 4sU-labeled RNA was submitted for sequencing and reads aligned to the mouse genome or MHV68 viral genome. Differential cellular gene expression was determined between mock and WT infected, mock and ∆HS infected, as well as differential viral gene expression between WT and ∆HS.

2015-07-23 | E-GEOD-70481 | biostudies-arrayexpress

Transcriptional analysis of an E2F gene signature as a biomarker of activity of the cyclin-dependent kinase inhibitor PHA-793887 in tumor and skin biopsies from a phase I clinical study

Project description:Microarray analysis of transcriptional changes in biopsies of solid tumors from patients treated in a Phase I study with the Cdk inhibitor PHA-793887. 7 patient biopsies pre- and post-treatment in dye swap. Tumors are of different origins, and are described in the accompanying manuscript.

2010-05-28 | E-GEOD-18553 | biostudies-arrayexpress

Gene expression data from yerba mate treated and non-treated cultured PBMCs activated with phytohemagglutinin

Project description:Yerba mate (YM) has been shown to have anti-inflammatory properties in several studies. However, this effect has been found mainly in obesity-related in inflammation. The aim of this work was to study the effect of YM in cultured peripheral blood mononuclear cells to see whether it has anti-inflammatory properties. We stimulated peripheral blood mononuclear cells in vitro with phitohemaglutinin in the presence of yerba mate and determined their activation measuring the the expression of CD25 by flow cytometry. We observed that YM treatment produced a dose-dependent reduction in PBMC activation (CD25 positive cells) when they were stimulated with PHA. This effect was also observed in T cells (CD3 positive) subpopulation. Microarray analysis revealed the differential expression of 128 genes in YM-treated cells. According to a protein-protein interaction database, these genes were highly connected and they are involved in inflammatory response. In summary, it was demonstrated that YM produces a reduction in the amount of activated cells under the stimulation of PHA. Therefore, it might be used in diseases with an inflammatory component. PBMC samples from a healthy individual were cultured and stimulated with phytohemagglutinin. Gene expression of yerba mate treated and non-treated cells was analyzed in duplicates, having in total four microarrays.

2016-08-15 | E-GEOD-84407 | biostudies-arrayexpress

Transcription profiling of human peripheral blood samples using different collection protocols - superseries of GSE12709, GSE12710

Project description:This SuperSeries is composed of the following subset Series:; GSE12709: Differential gene expression profiles are dependent upon method of peripheral blood RNA isolation (PHA); GSE12710: Differential gene expression profiles are dependent upon method of peripheral blood RNA isolation (direct, heparin) Experiment Overall Design: Refer to individual Series

2008-11-13 | E-GEOD-12711 | biostudies-arrayexpress

Unstressed HeLa cells and ELAVL1/HuR knock down conditions: polyA RNA-Seq, small RNA-Seq, and PAR-CLIP

Project description:Post-transcriptional gene regulation relies on hundreds of RNA binding proteins (RBPs) but the function of most RBPs is unknown. The human RBP HuR/ELAVL1 is a conserved mRNA stability regulator. We used PAR-CLIP, a method based on RNA-protein crosslinking, to identify transcriptome wide ~26,000 HuR binding sites. These sites were on average highly conserved, enriched for HuR binding motifs and mainly located in 3' untranslated regions. Surprisingly, many sites were intronic, implicating HuR in splicing. Upon HuR knock down, mRNA levels and protein synthesis of thousands of target genes was down regulated, validating functionality. HuR and miRNA binding sites tended to reside nearby but generally did not overlap. Additionally, HuR knock down triggered strong and specific up regulation of miR-7. In summary, we identified thousands of direct and functional HuR targets, found a human miRNA controlled by HuR, and propose a role for HuR in splicing. PolyA mRNA was extracted from anti HuR siRNA treated and mock transfected HeLa cells to identify changes in mRNA expression and splicing. 2x100 paired end sequencing was performed according to the protocol on the Illumina HiSeq. PARCLIP was performed as in Hafner et. Al 2010 but with an antibody against endogenous HuR (3A2, Santa Cruz, sc-5261) in unstressed HeLa cells. We used, independently, 4-thiouridine (4SU) and 6-thioguanosine (6SG) to assess a possible nucleotide bias. As our proteomics measurements required labeling of cells in a special medium we also performed PAR-CLIP on cells grown in SILAC medium. Altogether we performed three PAR-CLIP experiments: 4SU labeling in standard DMEM medium, 4SU labeling in SILAC medium (as a replicate) and 6SG labeling in SILAC medium. Small RNA was extracted from anti HuR siRNA treated and mock transfected HeLa cells to identify changes in mRNA expression. Sequencing was performed on Illumina GAII using the standard sRNA 36cycle protocol.

2011-06-29 | E-GEOD-29943 | biostudies-arrayexpress

Exon-expression profiling of CD4+ T cells derived from HTLV-1-infected individuals with or without malignancy

Project description:T-cell clones were obtained by limiting dilution culture of PBMC of HTLV-1 carriers. Exon expression profiling was performed using Affymetrix exon array (Affymetrix Human Exon 1.0 ST Array) according to the manufacturer's instructions. Gene version of CEL files 01 to 12 are presented in GSE46518. The main objectives were to assess whether transcriptional and post-transcriptional modifications associate with HTLV-1 infection in vivo. To this end, T-cell clones, infected or not by HTLV-1, were obtained by limiting dilution culture of PBMC derived from HTLV-1 carriers. Tumor cells derived from patients with an acute form ATLL. Exon expression profiles of cloned T-cells and ATLL cells was analyzed using Affymetrix exon array (Affymetrix Human Exon 1.0 ST Array) according to the manufacturer's instructions. Given that T-cell activation is known to modify alternative exon usage, microarray analysis was carried-out with unstimulated and PHA-stimulated CD4+ T cell clones.

2015-11-01 | E-GEOD-52244 | biostudies-arrayexpress

Immune response eQTL study in human neonatal myeloid cells and T cells

Project description:Gene expression microarray data in resting myeloid cells and CD4+ T cells, as well as in response to lipopolysaccharide (LPS) or phytohemagglutinin (PHA) stimulation for 24 hours, respectively. Myeloid cells and CD4+ T cells were purified from resting and stimulated peripheral blood mononuclear cells from cord blood samples of 152 individuals at birth. Raw array data and quantile normalised gene expression data are available. Full summary statistics of eQTL analysis and interaction tests for response eQTLs are available. Information such as sample attributes and experimental variables are available in the \\"Microarray_sample_info.txt\\".

2020-04-20 | E-MTAB-8977 | biostudies-arrayexpress

Temporal response of DCs to LPS stimulation: 4sU_sequencing

Project description:Regulation of RNA levels is critical for the response to external stimuli and determined through the interplay between RNA production, processing and degradation. Despite the centrality of these processes, most global studies of RNA regulation do not distinguish their separate contributions and relatively little is known about how they are temporally integrated. Here, we combine metabolic labeling of RNA with advanced RNA quantification assays and computational modeling to estimate RNA transcription and degradation during the response of immune dendritic cells (DCs) to pathogens, a critical and tightly regulated step in innate immunity. We find that transcription regulation plays a major role in shaping most temporal changes in RNA levels, but that changes in degradation rate are important for shaping sharp M-bM-^@M-^XpeakedM-bM-^@M-^Y responses. We find that transcription changes precede corresponding RNA changes by a small lag (15-30 min), which is shorter for induced than for repressed genes. Massively parallel sequencing of the entire RNA population M-bM-^@M-^S including non-polyadenylated transcripts M-bM-^@M-^S allows us to estimate RNA processing, and identify specific groups of transcripts, mostly cytokines and transcription factors, undergoing enhanced mRNA maturation. This suggests an additional role for splicing in regulating mRNA maturation. Our method provides a new quantitative approach to study key steps in the integrative process of RNA regulation. Sequencing of 4sU-labeled RNA taken from a 7 samples time-series (one sample every 1 hour) during the response of DCs to LPS stimulation. 4-thiouridine was added 45 minutes prior to sample collection. Data presented here for six timepoints: 0, 1, 3-6 hrs. 2hr timepoint not included.

2011-04-24 | E-GEOD-25432 | biostudies-arrayexpress

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