Global mapping of translation initiation sites in mammalian cells at single-nucleotide resolution
Global mapping of translation initiation sites in mammalian cells at single-nucleotide resolution
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DOI：
10.1073/pnas.1207846109
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发表时间：
2012-09-11
影响因子：
11.1
通讯作者：
Qian, Shu-Bing
中科院分区：
文献类型：
--
作者：
Lee, Sooncheol;Liu, Botao;Qian, Shu-Bing
理解基因表达中的翻译调控依赖于对整个转录组中翻译起始位点（TIS）的精确且全面的确定。最近开发的核糖体图谱技术能够进行全局翻译分析，提供了有关信使核糖核酸（mRNA）上核糖体位置和密度的大量信息。在此，我们提出一种方法，即全局翻译起始测序，同时应用核糖体E位点翻译抑制剂拉替米霉素和放线菌酮，以在全基因组范围内实现对起始和延伸事件的同时检测。这种方法以单核苷酸分辨率呈现了哺乳动物细胞中选择性翻译起始的情况。对翻译起始位点位置的系统性分析支持了翻译起始位点选择中的核糖体线性扫描机制。通过全局翻译起始测序确定的选择性翻译起始位点位置以及相关的开放阅读框在人类和小鼠细胞之间是保守的，这意味着选择性翻译具有生理意义。我们的研究建立了一个实用的平台，用于揭示转录组隐藏的编码潜力，并更好地理解翻译起始的复杂性。
Understanding translational control in gene expression relies on precise and comprehensive determination of translation initiation sites (TIS) across the entire transcriptome. The recently developed ribosome-profiling technique enables global translation analysis, providing a wealth of information about both the position and the density of ribosomes on mRNAs. Here we present an approach, global translation initiation sequencing, applying in parallel the ribosome E-site translation inhibitors lactimidomycin and cycloheximide to achieve simultaneous detection of both initiation and elongation events on a genome-wide scale. This approach provides a view of alternative translation initiation in mammalian cells with single-nucleotide resolution. Systemic analysis of TIS positions supports the ribosome linear-scanning mechanism in TIS selection. The alternative TIS positions and the associated ORFs identified by global translation initiation sequencing are conserved between human and mouse cells, implying physiological significance of alternative translation. Our study establishes a practical platform for uncovering the hidden coding potential of the transcriptome and offers a greater understanding of the complexity of translation initiation.