PPR-SMR protein SOT1 has RNA endonuclease activity
PPR-SMR protein SOT1 has RNA endonuclease activity
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DOI：
10.1073/pnas.1612460114
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发表时间：
2017-02
期刊：
影响因子：
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通讯作者：
Wen-jiao Zhou;Qingtao Lu;Qingwei Li;Lei Wang;Shunhua Ding;A. Zhang;Xiaogang Wen;Lixin Zhang
中科院分区：
文献类型：
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作者：
Wen-jiao Zhou;Qingtao Lu;Qingwei Li;Lei Wang;Shunhua Ding;A. Zhang;Xiaogang Wen;Lixin Zhang
我们的研究结果表明，类囊体形成的供应者1（SOT 1），拟南芥五肽重复（PPR）蛋白与一个小的MutS相关（SMR）结构域，具有内切酶活性。SOT 1的SMR部分通过PPR结构域特异性识别叶绿体23 S-4.5S rRNA前体5′端的13个核苷酸的RNA序列，进行23 S和4.5S rRNA的内切核酸成熟。我们的研究结果还表明，SOT 1可以被改造为识别和切割预测的RNA底物。我们的研究结果表明，SOT 1可以作为一种工具，在未来的RNA操作。已经进行了许多尝试来鉴定和工程化序列特异性RNA核酸内切酶，因为这些将允许有效的RNA操作。然而，迄今为止还没有描述以序列特异性方式识别RNA的天然RNA内切核酸酶。在这里，我们报告，类囊体形成的供应商1（SOT 1），拟南芥五肽重复（PPR）蛋白与一个小的MutS相关（SMR）域，具有RNA内切酶活性。我们发现SOT 1的SMR部分通过PPR结构域进行23 S和4.5S rRNA的核酸内切成熟，特异性识别叶绿体23 S-4.5S rRNA前体5′端的13个核苷酸的RNA序列。此外，我们成功地改造了SOT 1蛋白，改变了PPR基序，以识别和切割预测的RNA底物。我们的发现指出SOT 1是一种令人兴奋的RNA操纵工具。
Significance Our results demonstrate that SUPPRESSOR OF THYLAKOID FORMATION 1 (SOT1), an Arabidopsis pentatricopeptide repeat (PPR) protein with a small MutS-related (SMR) domain, has endonuclease activity. The SMR moiety of SOT1 performs the endonucleolytic maturation of 23S and 4.5S rRNA through the PPR domain specifically recognizing a 13-nucleotide RNA sequence in the 5′ end of the chloroplast 23S–4.5S rRNA precursor. Our results also show that SOT1 can be engineered to recognize and cleave a predicted RNA substrate. Our findings suggest that SOT1 could be used as a tool for RNA manipulation in the future. Numerous attempts have been made to identify and engineer sequence-specific RNA endonucleases, as these would allow for efficient RNA manipulation. However, no natural RNA endonuclease that recognizes RNA in a sequence-specific manner has been described to date. Here, we report that SUPPRESSOR OF THYLAKOID FORMATION 1 (SOT1), an Arabidopsis pentatricopeptide repeat (PPR) protein with a small MutS-related (SMR) domain, has RNA endonuclease activity. We show that the SMR moiety of SOT1 performs the endonucleolytic maturation of 23S and 4.5S rRNA through the PPR domain, specifically recognizing a 13-nucleotide RNA sequence in the 5′ end of the chloroplast 23S–4.5S rRNA precursor. In addition, we successfully engineered the SOT1 protein with altered PPR motifs to recognize and cleave a predicted RNA substrate. Our findings point to SOT1 as an exciting tool for RNA manipulation.