ATP is dispensable for both miRNA- and Smaug-mediated deadenylation reactions.
ATP is dispensable for both miRNA- and Smaug-mediated deadenylation reactions.
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DOI：
10.1261/rna.060764.117
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发表时间：
2017-06
期刊：
影响因子：
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通讯作者：
Tomari Y
中科院分区：
文献类型：
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作者：
Niinuma S;Tomari Y
MicroRNAs （miRNAs）以及rna结合蛋白Smaug招募CCR4-NOT死烯化酶复合物来缩短聚(A)尾部。基于ATP耗竭阻断死烯化反应的事实，认为miRNAs或Smaug诱导的死烯化需要ATP。然而，当分离时，CCR4-NOT复合体中的两种deadenylase本身都不需要ATP。因此，为什么核糖核蛋白复合物如mirna和Smaug的死基化需要ATP仍然是未知的。在这里，我们发现，在没有ATP再生系统的情况下，ATP在果蝇细胞裂解液中被迅速消耗成AMP，一种强的deadenylase抑制剂。重要的是，AMP的水解足以重新激活mirna或Smaug的死烯化，这表明AMP的积累，而不是ATP的消耗，导致了死烯化反应的抑制。我们的研究结果表明，ATP对于miRNAs或Smaug诱导的死基化是必不可少的，并强调在使用ATP耗尽方法时要谨慎。
MicroRNAs (miRNAs), as well as the RNA-binding protein Smaug, recruit the CCR4-NOT deadenylase complex for shortening of the poly(A) tail. It has been believed that ATP is required for deadenylation induced by miRNAs or Smaug, based on the fact that the deadenylation reaction is blocked by ATP depletion. However, when isolated, neither of the two deadenylases in the CCR4-NOT complex requires ATP by itself. Thus, it remains unknown why ATP is required for deadenylation by ribonucleoprotein complexes like miRNAs and Smaug. Herein we found that, in the absence of the ATP-regenerating system, ATP is rapidly consumed into AMP, a strong deadenylase inhibitor, in Drosophila cell lysate. Importantly, hydrolysis of AMP was sufficient to reactivate deadenylation by miRNAs or Smaug, suggesting that AMP accumulation, rather than ATP depletion, caused the inhibition of the deadenylation reaction. Our results indicate that ATP is dispensable for deadenylation induced by miRNAs or Smaug and emphasize caution in the use of ATP depletion methods.