Project description:We examined the possible effects of hypertonic stress on Arabidopsis translatome using polysome profiling. We found that the translatome is partly and rapidly reprogrammed in response to hypertonic stress, and such translatome reprogramming is DCP5-dependent.
Project description:au14-10_wd40 - effet of light on translatome of arabidopsis seeds during germination - Does light regulates germination via polysome association ? - At harvest seeds are dormant.They stay dormant if stored at -20°C.A.Th dormant seeds dont germinate at 25°C in darkness.Total RNA and polysomal RNA (polysomal fractions purified on sucrose gradients)were extracted from imbibed seeds for 20h at 22°C in darkness and light(3 biological replicates). Transcriptome and translatome are compared for light vs dark for 20h of imbibition. In silico comparison will allow to compare transcriptome and translatome for each type of sample.
Project description:RNA-binding proteins (RBPs) have essential functions during oocyte development.In order to explore the regulatory role of RNA-binding protein LSM14B in oocyte, we Identified the translatome of WT and Lsm14b KO GV-Stage Oocytes in mice via Scarce Sample Polysome Profiling (SSP-profiling).
Project description:Meiotic maturation of oocyte relies on pre-synthesized maternal mRNA, the translation of which is highly coordinated in space and time. Here, we provide a detailed polysome profiling protocol utilising small SW55Ti ultracentrifugation tubes with a volume of 5 ml. This newly optimized method, named Scarce Sample Polysome Profiling (SSP-profiling), is suitable for both scarce and conventional sample sizes and is compatible with downstream RNA-seq to identify polysome associated transcripts. Utilising SSP-profiling we have assayed the translatome of mouse oocytes at the onset of nuclear envelope breakdown (NEBD); a developmental stage that has thus far remained unexplored. Our analyses have identified 1778 transcripts with moderate to strong polysome occupancy, including abundantly represented mRNAs encoding mitochondrial and ribosomal proteins, proteoasomal components, glycolytic and amino acids synthetic enzymes, proteins involved in cytoskeleton organization plus RNA-binding and translation initiation factors. In addition to transcripts encoding known players of meiotic progression, we have also identified several mRNAs encoding proteins of unknown function. Polysome profiles generated using SSP-profiling were more than comparable to those developed using existing conventional approaches, being demonstrably superior in their resolution, reproducibility, versatility, speed of derivation and downstream protocol applicability.
Project description:cea03-02_translatome-cd - translatome-cd - Is there a change in the translation in Cadmium stress condition ? - Comparison between translated RNA (polysomes) and total RNA or non translated RNA (monosomes) in cadmium stress conditions. Keywords: transcribed vs translated,treated vs untreated comparison
Project description:In this study, we used CAGE followed by deep sequencing to globally profile the transcript 5’ isoforms in the translatome (i.e., polysome-associated RNA) and transcriptome (i.e., total RNA) of human HEK293 cells at single-nucleotide resolution. After removing low-quality sequencing reads, we obtained approximately 18 million and 14 million CAGE tags respectively for the translatome and transcriptome of HEK293 cells. These tags were then mapped to the human genome (assembly GRCh37) using bowtie with two mismatches allowed. Tags mapped to rRNA were less than 17.9% for translatome and 7.5% for transcriptome, indicating high quality of the two CAGE libraries. By comparing CAGE tags between HEK293's translatome and transcriptome, we revealed selective usage of the TSS-derived 5’ ends by polysome.
Project description:Heat stress is one of the most prominent and deleterious environmental threads affecting plant growth and development. Upon high temperatures, plants launch specialized gene expression programs that promote stress protection and survival. These programs involve global and specific changes at the transcriptional and translational levels. However the coordination of these processes and their specific role in the establishment of the heat stress response is not fully elucidated. In this report, we have carried out a genome-wide analysis to simultaneously monitor the individual changes in the transcriptional and translational mRNA levels of Arabidopsis thaliana seedlings after the exposure to a heat shock stress. Our results demonstrated that, superimposed to transcription, translation exerts a wide but dual regulation of gene expression. For the majority of mRNAs, translation is severely repressed, causing a decreased of 50% of the association of the bulk of mRNAs to polysomes. However, some relevant mRNAs involved in different aspects of homeostasis maintenance follow a differential pattern of translation. Analysis of the sequence of the differentially translated mRNAs unravels that some features, like the 5M-BM-4UTR G+C content and the cDNA length, may take part in the discrimination mechanisms for mRNA polysome loading. Among the differential translated genes stand out master regulators of the stress response, highlighting the main role of translation in the early establishment of physiological response of plants to elevated temperatures. In total 8 ATH1 Affymetrix GeneChips were hybridized with all combinations of two factors: total mRNA/polysome-bound-RNA; 22M-BM-:C/38M-BM-:C. Two biological replicates per sample type were performed.
Project description:FXR1 overexpression in THP1 cells causes changes in polysome associated mRNAs. This leads to a distinct translatome in the cell that contributes to chemo- and immune- survival