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The S. cerevisiae m6A reader Pho92 promotes timely meiotic recombination by controlling key methylated transcripts [CRAC]

ABSTRACT: We performed CRAC (UV-crosslinking and analysis of cDNA) to identify Ime4-dependent mRNA targets of Pho92 during meiosis at 5 hr in SPO.

ORGANISM(S): Saccharomyces cerevisiae

PROVIDER: GSE200088 | GEO | 2022/07/08

REPOSITORIES: GEO

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The S. cerevisiae m6A reader Pho92 promotes timely meiotic recombination by controlling key methylated transcripts [RNA-seq]

Project description:We performed high-throughput mRNA sequencing during a meiotic time course (0, 2, 3, 5, 6, 7 and 9 hr) in WT, pho92 mutant and ime4 mutant cells.

2022-07-08 | GSE200087 | GEO

The S. cerevisiae m6A reader Pho92 promotes timely meiotic recombination by controlling key methylated transcripts [CRAC]

Project description:The S. cerevisiae m6A reader Pho92 promotes timely meiotic recombination by controlling key methylated transcripts [CRAC]

| PRJNA822916 | ENA

The S. cerevisiae m6A reader Pho92 promotes timely meiotic recombination by controlling key methylated transcripts.

Project description:This SuperSeries is composed of the SubSeries listed below.

2022-07-08 | GSE200089 | GEO

The S. cerevisiae m6A reader Pho92 promotes timely meiotic recombination by controlling key methylated transcripts.

Project description:The S. cerevisiae m6A reader Pho92 promotes timely meiotic recombination by controlling key methylated transcripts.

| PRJNA822912 | ENA

The S. cerevisiae m6A reader Pho92 promotes timely meiotic recombination by controlling key methylated transcripts [RNA-seq]

Project description:The S. cerevisiae m6A reader Pho92 promotes timely meiotic recombination by controlling key methylated transcripts [RNA-seq]

| PRJNA822917 | ENA

m6A reader Pho92 is recruited co-transcriptionally and couples translation efficacy to mRNA decay to promote meiotic fitness in yeast [iCLIP_miCLIP-seq]

Project description:During meiosis, in Saccharomyces cerevisiae, N6-methyladenosine (m6A) modified transcripts are induced, of which the function is unknown. Here, we uncover the role of the m6A reader Pho92. Cross-linking immunoprecipitation (CLIP) revealed that Pho92 associates with meiotic mRNAs in both m6A dependent and independent manner. Incidentally, Pho92 resides in the nucleus during early meiosis and associates with nascent RNAs, which is mediated through its interaction with Paf1C. Transcripts bound by Pho92 show elevated translational efficiency while cells lacking Pho92 display a small, but notable, increase in mRNA levels but not in protein levels, suggesting role of Pho92 in translation and decay. We show that Pho92 associates with ribosomes where it promotes the decay of m6A modified transcripts, contingent on active translation and the CCR4-NOT complex. We propose that m6A reader Pho92 is loaded co-transcriptionally to promote translation and subsequent decay fate of m6A modified transcripts, which ensures gamete fitness.

2022-12-06 | GSE193291 | GEO

m6A reader Pho92 is recruited co-transcriptionally and couples translation efficacy to mRNA decay to promote meiotic fitness in yeast [Polysome]

Project description:N6-methyladenosine (m6A) RNA modification impacts mRNA fate primarily via reader proteins, which dictate processes in development, stress, and disease. Yet little is known about m6A function in Saccharomyces cerevisiae, which occurs solely during early meiosis. Here we perform a multifaceted analysis of the m6A reader protein Pho92/Mrb1. Cross-linking immunoprecipitation analysis reveals that Pho92 associates with the 3’end of meiotic mRNAs in both an m6A-dependent and independent manner. Within cells, Pho92 transitions from the nucleus to the cytoplasm, and associates with translating ribosomes. In the nucleus Pho92 associates with target loci through its interaction with transcriptional elongator Paf1C. Functionally, we show that Pho92 promotes and links protein synthesis to mRNA decay. As such, the Pho92-mediated m6A-mRNA decay is contingent on active translation and the CCR4-NOT complex. We propose that the m6A reader Pho92 is loaded co-transcriptionally to facilitate correct translation and subsequent decay of m6A modified transcripts, and thereby promotes meiosis.

2022-12-06 | GSE193560 | GEO

m6A reader Pho92 is recruited co-transcriptionally and couples translation efficacy to mRNA decay to promote meiotic fitness in yeast [ime1D_ndt80D]

2022-12-06 | GSE193558 | GEO

m6A reader Pho92 is recruited co-transcriptionally and couples translation efficacy to mRNA decay to promote meiotic fitness in yeast [pho92D_ime4D]

2022-12-06 | GSE193559 | GEO

Characterization of A Novel CRAC Inhibitor that Potently Blocks Human T cell Activation and Effector Functions

Project description:Store operated calcium entry (SOCE) downstream of T cell receptor (TCR) activation in T lymphocytes has been shown to be mediated mainly through the Calcium Release Activated Calcium (CRAC) channel. Here, we compared the effects of a novel, potent and selective CRAC inhibitor, 2,6-Difluoro-N-{5-[4-methyl-1-(5-methyl-thiazol-2-yl)-1,2,5,6-tetrahydro-pyridin-3-yl]-pyrazin-2-yl}-benzamide (RO2959), on T cell effector functions with that of a previously reported CRAC channel inhibitor, YM-58483, and a calcineurin inhibitor Cyclosporin A (CsA). Using both electrophysiological and calcium-based fluorescence measurements, we showed that RO2959 is a potent SOCE inhibitor that blocked an IP3-dependent current in CRAC-expressing RBL-2H3 cells and CHO cells stably expressing human Orai1 and Stim1, as well as SOCE in human primary CD4+ T cells triggered by either TCR stimulation or thapsigargin treatment. Furthermore, we demonstrated that RO2959 completely inhibited cytokine production as well as T cell proliferation mediated by TCR stimulation or MLR (Mixed Lymphocyte Reaction). Lastly, we showed by gene expression array analysis that RO2959 potently blocked TCR triggered gene expression and T cell functional pathways similar to CsA and FK506. Thus, both from a functional and transcriptional level, our data provide evidence that RO2959 is a novel and selective CRAC inhibitor that potently inhibits human T cell functions. PBMC from healthy donors (n=4) were stimulated with anti-CD3/CD28 in the presence or absence of CRAC inhibitor, CsA or FK506 for 24 hrs

2013-01-31 | E-GEOD-43210 | biostudies-arrayexpress

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