Department of Microbiology and Molecular Genetics, Michigan State University, East Lansing, MI 48824, USA, Department of Computer Science and Engineering, Michigan State University, East Lansing, MI 48824, USA and Department of Biochemistry and Molecular Biology, Michigan State University, East Lansing, MI 48824, USA.
Nucleic Acids Res. 2014 Feb;42(3):1873-86. doi: 10.1093/nar/gkt973. Epub 2013 Oct 29.
One of the most striking examples of small RNA regulation of gene expression is the process of RNA editing in the mitochondria of trypanosomes. In these parasites, RNA editing involves extensive uridylate insertions and deletions within most of the mitochondrial messenger RNAs (mRNAs). Over 1200 small guide RNAs (gRNAs) are predicted to be responsible for directing the sequence changes that create start and stop codons, correct frameshifts and for many of the mRNAs generate most of the open reading frame. In addition, alternative editing creates the opportunity for unprecedented protein diversity. In Trypanosoma brucei, the vast majority of gRNAs are transcribed from minicircles, which are approximately one kilobase in size, and encode between three and four gRNAs. The large number (5000-10,000) and their concatenated structure make them difficult to sequence. To identify the complete set of gRNAs necessary for mRNA editing in T. brucei, we used Illumina deep sequencing of purified gRNAs from the procyclic stage. We report a near complete set of gRNAs needed to direct the editing of the mRNAs.
小 RNA 对基因表达调控的一个最显著例子是原生动物线粒体中的 RNA 编辑过程。在这些寄生虫中，RNA 编辑涉及大多数线粒体信使 RNA（mRNA）中的广泛尿嘧啶插入和缺失。预计有超过 1200 个小指导 RNA（gRNA）负责指导产生起始和终止密码子、纠正移码以及为大多数 mRNA 生成大部分开放阅读框的序列变化。此外，替代编辑为前所未有的蛋白质多样性创造了机会。在布氏锥虫中，绝大多数 gRNA 是从小环中转录而来的，小环大小约为 1000 个碱基，编码 3 到 4 个 gRNA。数量众多（5000-10000）及其串联结构使得它们难以测序。为了鉴定在 T. brucei 中指导 mRNA 编辑所需的完整 gRNA 集，我们使用 Illumina 对来自前鞭毛体阶段的纯化 gRNA 进行了深度测序。我们报告了一组近乎完整的 gRNA，这些 gRNA 用于指导 mRNA 的编辑。
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