Global view on the metabolism of RNA poly(A) tails in yeast Saccharomyces cerevisiae.
Global view on the metabolism of RNA poly(A) tails in yeast Saccharomyces cerevisiae.
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DOI：
10.1038/s41467-021-25251-w
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发表时间：
2021-08-16
影响因子：
16.6
通讯作者：
Dziembowski A
中科院分区：
文献类型：
--
作者：
Tudek A;Krawczyk PS;Mroczek S;Tomecki R;Turtola M;Matylla-Kulińska K;Jensen TH;Dziembowski A
聚腺苷尾 (poly[A]-tail) 是真核信使 RNA (mRNA) 和非编码 RNA (ncRNA) 的通用修饰。在芽殖酵母中，Pap1 合成的 mRNA 聚腺苷酸尾增强了输出和翻译，而 Trf4/5 介导的 ncRNA 聚腺苷酸化则促进了外泌体的降解。使用直接 RNA 测序，我们破译了所有相关核酸外切酶、去腺苷酸酶和聚腺苷酸聚合酶缺陷的酵母中聚腺苷酸尾部动力学的程度。主要的 ncRNA Poly(A) 尾部长度为 20-60 个腺苷。新转录的 mRNA 的 Poly(A) 尾部平均长度为 50 个腺苷，上限为 200 个。Trf5 辅助的核外泌体和细胞质去腺苷酶进行核酸外切，将尾部平均修剪为 40 个腺苷。令人惊讶的是，PAN2/3 和 CCR4-NOT 去腺苷酸酶复合物具有大量主要由表达水平定义的非重叠底物。最后，我们证明 mRNA Poly(A) 尾长对生长条件（例如热量和营养缺乏）有强烈反应。 RNA 聚腺苷尾对于 mRNA 的输出、翻译和稳定性很重要，并在非编码 RNA 生物合成中发挥作用。在这里，作者通过直接 RNA 测序测量了酵母 Poly(A) 尾长，揭示了其在酵母核酸外切酶、去腺苷酸酶和 Poly(A) 聚合酶突变体中的动态。
The polyadenosine tail (poly[A]-tail) is a universal modification of eukaryotic messenger RNAs (mRNAs) and non-coding RNAs (ncRNAs). In budding yeast, Pap1-synthesized mRNA poly(A) tails enhance export and translation, whereas Trf4/5-mediated polyadenylation of ncRNAs facilitates degradation by the exosome. Using direct RNA sequencing, we decipher the extent of poly(A) tail dynamics in yeast defective in all relevant exonucleases, deadenylases, and poly(A) polymerases. Predominantly ncRNA poly(A) tails are 20-60 adenosines long. Poly(A) tails of newly transcribed mRNAs are 50 adenosine long on average, with an upper limit of 200. Exonucleolysis by Trf5-assisted nuclear exosome and cytoplasmic deadenylases trim the tails to 40 adenosines on average. Surprisingly, PAN2/3 and CCR4-NOT deadenylase complexes have a large pool of non-overlapping substrates mainly defined by expression level. Finally, we demonstrate that mRNA poly(A) tail length strongly responds to growth conditions, such as heat and nutrient deprivation. RNA polyadenosine tails are important for the export, translation and stability of mRNAs and play a role in non-coding RNA biogenesis. Here the authors measure yeast poly(A) tail lengths by direct RNA sequencing, revealing its dynamics in yeast exonuclease, deadenylase and poly(A) polymerase mutants.
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