Jiao Yuling, Riechmann José Luis, Meyerowitz Elliot M
California Institute of Technology, Division of Biology 156-29, Pasadena, California 91125, USA.
Plant Cell. 2008 Oct;20(10):2571-85. doi: 10.1105/tpc.108.062786. Epub 2008 Oct 24.
The composition of the transcriptome is determined by a balance between mRNA synthesis and degradation. An important route for mRNA degradation produces uncapped mRNAs, and this decay process can be initiated by decapping enzymes, endonucleases, and small RNAs. Although uncapped mRNAs are an important intermediate for mRNA decay, their identity and abundance have never been studied on a large scale until recently. Here, we present an experimental method for transcriptome-wide profiling of uncapped mRNAs that can be used in any eukaryotic system. We applied the method to study the prevalence of uncapped transcripts during the early stages of Arabidopsis thaliana flower development. Uncapped transcripts were identified for the majority of expressed genes, although at different levels. By comparing uncapped RNA levels with steady state overall transcript levels, our study provides evidence for widespread mRNA degradation control in numerous biological processes involving genes of varied molecular functions, implying that uncapped mRNA levels are dynamically regulated. Sequence analyses identified structural features of transcripts and cis-elements that were associated with different levels of uncapping. These transcriptome-wide profiles of uncapped mRNAs will aid in illuminating new regulatory mechanisms of eukaryotic transcriptional networks.
转录组的组成由mRNA合成与降解之间的平衡决定。mRNA降解的一条重要途径会产生无帽mRNA,这种衰变过程可由去帽酶、核酸内切酶和小RNA启动。尽管无帽mRNA是mRNA衰变的重要中间体,但直到最近才对其身份和丰度进行大规模研究。在这里,我们提出了一种用于对无帽mRNA进行全转录组分析的实验方法,该方法可用于任何真核系统。我们应用该方法研究了拟南芥花发育早期无帽转录本的普遍性。尽管表达水平不同,但大多数表达基因都鉴定出了无帽转录本。通过将无帽RNA水平与稳态总体转录本水平进行比较,我们的研究为涉及多种分子功能基因的众多生物过程中广泛的mRNA降解控制提供了证据,这意味着无帽mRNA水平受到动态调节。序列分析确定了与不同去帽水平相关的转录本结构特征和顺式元件。这些无帽mRNA的全转录组图谱将有助于阐明真核转录网络的新调控机制。