Yeast Dxo1 is required for 25S rRNA maturation and acts as a transcriptome-wide distributive exonuclease.
Yeast Dxo1 is required for 25S rRNA maturation and acts as a transcriptome-wide distributive exonuclease.
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DOI：
10.1261/rna.078952.121
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发表时间：
2022-05
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Dxo1/Rai1/DXO家族的脱帽酶和核酸外切酶可以在体外催化从RNA中去除化学上不同的5 '端。具体来说，这些酶对具有标准7mGpppN帽的RNA的作用很差，但更倾向于具有三磷酸、单磷酸、羟基或非常规帽的RNA。在每种情况下，这些酶产生具有5 ‘单磷酸的RNA，然后被认为被Rat1/Xrn1 5 ’外核糖核酸酶进一步降解。对于大多数DXO /Rai1/DXO家族成员，尚不清楚这些活性中哪一个在体内最重要。在这里，我们描述了细胞质家族成员酵母Dxo1的体内功能。利用5 '单磷酸末端的RNA-seq，我们发现Dxo1可以作为一种分布性核酸外切酶，从核酸内切酶或脱盖产物中去除一些核苷酸。我们还表明，Dxo1是25S rRNA最后的5 '端加工所必需的，这是Dxo1的主要作用。虽然Dxo1/Rai1/DXO成员预计作用于Rat1/Xrn1的上游，但在25S rRNA加工中，这一顺序相反，Dxo1作用于Rat1的下游。这种从过程性到分布性的外切酶的转换可能是RNA末端精确成熟过程中的普遍现象。
The Dxo1/Rai1/DXO family of decapping and exonuclease enzymes can catalyze the in vitro removal of chemically diverse 5′ ends from RNA. Specifically, these enzymes act poorly on RNAs with a canonical 7mGpppN cap, but instead prefer RNAs with a triphosphate, monophosphate, hydroxyl, or nonconventional cap. In each case, these enzymes generate an RNA with a 5′ monophosphate, which is then thought to be further degraded by Rat1/Xrn1 5′ exoribonucleases. For most Dxo1/Rai1/DXO family members, it is not known which of these activities is most important in vivo. Here we describe the in vivo function of the poorly characterized cytoplasmic family member, yeast Dxo1. Using RNA-seq of 5′ monophosphate ends, we show that Dxo1 can act as a distributive exonuclease, removing a few nucleotides from endonuclease or decapping products. We also show that Dxo1 is required for the final 5′ end processing of 25S rRNA, and that this is the primary role of Dxo1. While Dxo1/Rai1/DXO members were expected to act upstream of Rat1/Xrn1, this order is reversed in 25S rRNA processing, with Dxo1 acting downstream from Rat1. Such a hand-off from a processive to a distributive exonuclease may be a general phenomenon in the precise maturation of RNA ends.
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