U2 snRNA is inducibly pseudouridylated at novel sites by Pus7p and snR81 RNP

U2 snRNA is inducibly pseudouridylated at novel sites by Pus7p and snR81 RNP
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DOI:
10.1038/emboj.2010.316
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发表时间:
2011-01-05
期刊:
影响因子:
11.4
通讯作者:
Yu, Yi-Tao
Yu, Yi-Tao
中科院分区:
生物学1区
文献类型:
--
作者:
Wu, Guowei;Xiao, Mu;Yu, Yi-Tao

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RNA 中鉴定出的所有假尿苷均被视为组成型修饰。在这里,我们证明酿酒酵母 U2 snRNA 的假尿苷化可以被条件诱导。虽然在正常条件下仅在 112 中检测到 Psi 35、Psi 42 和 Psi 44,但营养缺乏会导致位置 56 和 93 处出现额外的假尿苷化。位置 56 处的假尿苷化也可由热休克诱导。详细分析表明,Pus7p(已知修饰 112 位点 35 和 tRNA 修饰 13 位点的单多肽假尿苷酸酶)催化 Psi 56 的形成,而 snR81 RNP(已知修饰位点 42 的 112 snRNA 和位点 1051 的 25S rRNA 的框 H/ACA RNP)催化 Psi 93 的形成。通过诱变,我们证明了诱导性可归因于不完美的底物序列。通过将 Psi 93 引入对数期细胞,我们进一步表明 Psi 93 在前 mRNA 剪接中发挥作用。因此,我们的结果首次证明,RNA 的假尿苷化可以在不完美序列的位点诱导,并且 Pus7p 和 snR81 RNP 可以催化组成型和诱导型假尿苷化。 EMBO 杂志 (2011) 30, 79-89。 doi:10.1038/ emboj.2010.316;在线发表 2010 年 12 月 3 日 主题类别:RNA
All pseudouridines identified in RNA are considered constitutive modifications. Here, we demonstrate that pseudouridylation of Saccharomyces cerevisiae U2 snRNA can be conditionally induced. While only Psi 35, Psi 42 and Psi 44 are detected in 112 under normal conditions, nutrient deprivation leads to additional pseudouridylation at positions 56 and 93. Pseudouridylation at position 56 can also be induced by heat shock. Detailed analyses have shown that Pus7p, a single polypeptide pseudouridylase known to modify 112 at position 35 and tRNA at position 13, catalyses Psi 56 formation, and that snR81 RNP, a box H/ACA RNP known to modify 112 snRNA at position 42 and 25S rRNA at position 1051, catalyses Psi 93 formation. Using mutagenesis, we have demonstrated that the inducibility can be attributed to the imperfect substrate sequences. By introducing Psi 93 into log-phase cells, we further show that Psi 93 has a role in pre-mRNA splicing. Our results thus demonstrate for the first time that pseudouridylation of RNA can be induced at sites of imperfect sequences, and that Pus7p and snR81 RNP can catalyse both constitutive and inducible pseudouridylation. The EMBO Journal (2011) 30, 79-89. doi:10.1038/ emboj.2010.316; Published online 3 December 2010 Subject Categories: RNA