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Novel RNA partners of the Mei2 RNA binding protein during early meiosis in Schizosaccharomyces pombe

ABSTRACT: We used CLIP-Seq to determine the RNAs bound specifically to RNA binding protein Mei2 in early meiosis in fission yeast. We added a TAP tag to the C-terminal ends of two meiotic RNA binding proteins, Mei2 and Msa1. We used an untagged fission yeast strain as a negative control. These strains were nitrogen starved and allowed to progress into meiosis, after which they were harvested, lyzed and crosslinking immunoprecipitaion was performed. The RNAs purified after CLIP were sequenced

ORGANISM(S): Schizosaccharomyces pombe

PROVIDER: GSE112117 | GEO | 2019/01/01

REPOSITORIES: GEO

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Novel RNA partners of the Mei2 RNA binding protein during early meiosis in Schizosaccharomyces pombe

Project description:We used RIP-CHIP to determine the RNAs bound specifically to RNA binding protein Mei2 in early meiosis in fission yeast.

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A functional genomics screen identifies genes required for Schizosaccharomyces pombe meiosis

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Distinctive responses to nitrogen starvation in the dominant active mutants of the fission yeast Rheb GTPase.

Project description:Rheb, a ras-like small GTPase conserved from human to yeast, controls Tor kinase and plays a central role in regulation of cell growth depending on extracellular conditions. Fission yeast Rheb regulates amino acid uptake as well as response to nitrogen starvation. In this study we generated two mutants of Rheb, rhb1-DA4 and rhb1-DA8, and characterized them genetically. V17A mutation within the G1 box defined for the ras-like GTPases was responsible for rhb1-DA4, and Q52R I76F within the switch II domain for rhb1-DA8. In fission yeast, two events, induction of a meiosis initiating gene mei2+ and cell division without cell growth, are a typical response to nitrogen starvation. Under nitrogen-rich conditions, Rheb stimulates Tor kinase, which, in turn, suppresses the response to nitrogen starvation. While amino acid uptake was prevented by both rhb1-DA4 and rhb1-DA8 in a dominant fashion, the response to nitrogen starvation was prevented only by rhb1-DA4. rhb1-DA8 thereby allowed genetic dissection of the Rheb-dependent signaling cascade. We postulate that Rheb in fission may have two downstream elements, Tor kinase for regulation of the response to nitrogen starvation and the other element for regulation of amino acid uptake. Gene expression profile under nitrogen starvation in fission yeast. Type of experiment: Comparing between vegetatively-growing control cells and cells 3 hrs after nitrogen starvation. Experimental factor: Gene expression profile after nitrogen starvation in the rhb1-D4 or rhb1-D8 cells. Quality control steps taken: All experiments were repeated more than twice except for the tsc2D which was previously reported. Keywards: Nitrogen starvation, rhb1-D4, rhb1-D8

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