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Nonsense-mediated decay does not occur within the yeast nucleus

机译:无义介导的衰变不会在酵母核内发生

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摘要

Nonsense-mediated decay (NMD) is a eukaryotic regulatory process that degrades mRNAs with premature termination codons (PTCs). Although NMD is a translation-dependent process, there is evidence from mammalian systems that PTC recognition and mRNA degradation takes place in association with nuclei. Consistent with this notion, degradation of mammalian PTC-containing mRNAs occurs when they are bound by the cap binding complex (CBC) during a “pioneer” round of translation. Moreover, there are reports indicating that a PTC can trigger other nuclear events such as alternative splicing, abnormal 3′ end processing, and accumulation of pre-mRNA at transcription sites. To examine whether a PTC can elicit similar nuclear events in yeast, we used RNA export-defective mutants to sequester mRNAs within nuclei. The results indicate that nuclear PTC-containing yeast RNAs are NMD insensitive. We also observed by fluorescent in situ hybridization that there was no PTC effect on mRNA accumulated at the site of transcription. Finally, we show that yeast NMD occurs minimally if at all on CBC-bound transcripts, arguing against a CBC-mediated pioneer round of translation in yeast. The data taken together indicate that there are no direct consequences of a PTC within the yeast nucleus.
机译:无意义介导的衰变(NMD)是一种真核调节过程,可通过过早终止密码子(PTC)降解mRNA。尽管NMD是依赖翻译的过程,但哺乳动物系统的证据表明PTC识别和mRNA降解与细胞核有关。与该概念一致,当在“开拓者”轮的翻译过程中,包含PTC的哺乳动物mRNA被帽结合结合物(CBC)结合时,就会发生降解。此外,有报道表明PTC可以触发其他核事件,例如选择性剪接,异常3'末端加工以及pre-mRNA在转录位点的积累。为了检查PTC是否可以在酵母中引发相似的核事件,我们使用了RNA出口缺陷型突变体将核仁中的mRNA隔离。结果表明,含核PTC的酵母RNA对NMD不敏感。我们还通过荧光原位杂交观察到,PTC对转录位点处积累的mRNA没有影响。最后,我们证明酵母NMD如果在CBC结合的转录本上根本没有发生,则与酵母中CBC介导的先驱回合有关。汇总的数据表明,PTC在酵母核内没有直接影响。

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