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The Fission Yeast RNA-Binding Protein Meu5 Is Involved in Outer Forespore Membrane Breakdown during Spore Formation

机译:裂变酵母RNA结合蛋白MEU5涉及孢子形成期间的外额外介质膜分解

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摘要

In Schizosaccharomyces pombe, the spore wall confers strong resistance against external stress. During meiosis II, the double-layered intracellular forespore membrane (FSM) forms de novo and encapsulates the nucleus. Eventually, the inner FSM layer becomes the plasma membrane of the spore, while the outer layer breaks down. However, the molecular mechanism and biological significance of this membrane breakdown remain unknown. Here, by genetic investigation of an S. pombe mutant (E22) with normal prespore formation but abnormal spores, we showed that Meu5, an RNA-binding protein known to bind to and stabilize more than 80 transcripts, is involved in this process. We confirmed that the E22 mutant does not produce Meu5 protein, while overexpression of meu5+ in E22 restores the sporulation defect. Furthermore, electron microscopy revealed that the outer membrane of the FSM persisted in meu5∆ spores. Investigation of the target genes of meu5+ showed that a mutant of cyc1+ encoding cytochrome c also showed a severe defect in outer FSM breakdown. Lastly, we determined that outer FSM breakdown occurs coincident with or after formation of the outermost Isp3 layer of the spore wall. Collectively, our data provide novel insights into the molecular mechanism of spore formation.
机译:在Schizosaccharomyces Pombe中,孢子壁赋予了强烈的对抗外部压力的抗性。在减数分裂II期间,双层细胞内患者膜(FSM)形成De Novo并包封核。最终,内部FSM层成为孢子的质膜,而外层断裂。然而,这种膜击穿的分子机制和生物学意义仍然是未知的。这里,通过具有正常的预参数形成但孢子异常的S.Pombe突变体(E22)的遗传调查,我们表明MEU5,已知的RNA结合蛋白结合并稳定大于80种转录物,参与该方法。我们证实,E22突变体不会产生MEU5蛋白,而E22中MEU5 +的过​​度表达恢复孢子缺陷。此外,电子显微镜显示,FSM的外膜持续在MEU5δ孢子中。 MEU5 +靶基因的研究表明,CYC1 +编码细胞色素C的突变体也显示出外部FSM击穿的严重缺陷。最后,我们确定了外部FSM击穿发生在孢子墙的最外面的ISP3层的形成中或在形成之后。集体,我们的数据为孢子形成的分子机制提供了新的洞察力。

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