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Rpl22对于酿酒酵母中IME1 mRNA的翻译和减数分裂诱导是必需的。

Rpl22 is required for IME1 mRNA translation and meiotic induction in S. cerevisiae.

作者信息

Kim Stephen J, Strich Randy

机构信息

Department of Molecular Biology, Rowan University School of Osteopathic Medicine, Two Medical Center Dr., Stratford, NJ 08055 USA.

出版信息

Cell Div. 2016 Jul 29;11:10. doi: 10.1186/s13008-016-0024-3. eCollection 2016.

Abstract

BACKGROUND

The transition from mitotic cell division to meiotic development in S. cerevisiae requires induction of a transient transcription program that is initiated by Ime1-dependent destruction of the repressor Ume6. Although IME1 mRNA is observed in vegetative cultures, Ime1 protein is not suggesting the presence of a regulatory system restricting translation to meiotic cells.

RESULTS

This study demonstrates that IME1 mRNA translation requires Rpl22A and Rpl22B, eukaryotic-specific ribosomal protein paralogs of the 60S large subunit. In the absence of Rpl22 function, IME1 mRNA synthesis is normal in cultures induced to enter meiosis. However, Ime1 protein production is reduced and the Ume6 repressor is not destroyed in rpl22 mutant cells preventing early meiotic gene induction resulting in a pre-meiosis I arrest. This role for Rpl22 is not a general consequence of mutating non-essential large ribosomal proteins as strains lacking Rpl29 or Rpl39 execute meiosis with nearly wild-type efficiencies. Several results indicate that Rpl22 functions by enhancing IME1 mRNA translation. First, the Ime1 protein synthesized in rpl22 mutant cells demonstrates the same turnover rate as in wild-type cultures. In addition, IME1 transcript is found in polysome fractions isolated from rpl22 mutant cells indicating that mRNA nuclear export and ribosome association occurs. Finally, deleting the unusually long 5'UTR restores Ime1 levels and early meiotic gene transcription in rpl22 mutants suggesting that Rpl22 enhances translation through this element. Polysome profiles revealed that under conditions of high translational output, Rpl22 maintains high free 60S subunit levels thus preventing halfmer formation, a translation species indicative of mRNAs bound by an unpaired 40S subunit. In addition to meiosis, Rpl22 is also required for invasive and pseudohyphal growth.

CONCLUSIONS

These findings indicate that Rpl22A and Rpl22B are required to selectively translate IME1 mRNA that is required for meiotic induction and subsequent gametogenesis. In addition, our results imply a more general role for Rpl22 in cell fate switches responding to environmental nitrogen signals.

摘要

背景

酿酒酵母从有丝分裂细胞分裂向减数分裂发育的转变需要诱导一个短暂的转录程序,该程序由Ime1依赖的阻遏物Ume6的破坏启动。尽管在营养培养物中观察到IME1 mRNA,但未观察到Ime1蛋白,这表明存在一个将翻译限制在减数分裂细胞中的调节系统。

结果

本研究表明,IME1 mRNA翻译需要Rpl22A和Rpl22B,它们是60S大亚基的真核特异性核糖体蛋白旁系同源物。在缺乏Rpl22功能的情况下,诱导进入减数分裂的培养物中IME1 mRNA合成正常。然而,rpl22突变细胞中Ime1蛋白的产生减少,Ume6阻遏物未被破坏,从而阻止了减数分裂早期基因的诱导,导致减数分裂I前期停滞。Rpl22的这一作用不是突变非必需的大核糖体蛋白的一般结果,因为缺乏Rpl29或Rpl39的菌株以接近野生型的效率进行减数分裂。几个结果表明,Rpl22通过增强IME1 mRNA翻译发挥作用。首先,在rpl22突变细胞中合成的Ime1蛋白显示出与野生型培养物中相同的周转率。此外,在从rpl22突变细胞中分离的多核糖体组分中发现了IME1转录本,表明mRNA的核输出和核糖体结合发生。最后,删除异常长的5'UTR可恢复rpl22突变体中的Ime1水平和减数分裂早期基因转录,这表明Rpl22通过该元件增强翻译。多核糖体图谱显示,在高翻译输出条件下,Rpl22维持高游离60S亚基水平,从而防止半聚体形成,半聚体是一种由未配对的40S亚基结合的mRNA的翻译物种。除了减数分裂,Rpl22也是侵袭性和假菌丝生长所必需的。

结论

这些发现表明,Rpl22A和Rpl22B是选择性翻译减数分裂诱导和随后配子发生所需的IME1 mRNA所必需的。此外,我们的结果暗示Rpl22在响应环境氮信号的细胞命运转换中具有更广泛的作用。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/10c1/4966820/f8aede15276d/13008_2016_24_Fig1_HTML.jpg

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