Topology of the human and mouse m6A RNA methylomes revealed by m6A-seq
Topology of the human and mouse m6A RNA methylomes revealed by m6A-seq
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DOI：
10.1038/nature11112
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发表时间：
2012-05-10
期刊：
影响因子：
64.8
通讯作者：
Rechavi, Gideon
中科院分区：
文献类型：
--
作者：
Dominissini, Dan;Moshitch-Moshkovitz, Sharon;Rechavi, Gideon
已知RNA的多种编码、结构和催化功能是由大量修饰组成的，但在很大程度上仍是未知领域。尽管生物化学研究表明N-6-甲基腺苷（m（6）A）是信使RNA中最普遍的内部修饰，但由于缺乏可靠的分析方法，对其分布和功能的深入研究受到阻碍。在这里，我们使用一种基于抗体介导的捕获和大规模平行测序的新方法m（6）A-seq，以全转录组的方式呈现人类和小鼠m（6）A修饰景观。我们在7,000多个人类基因的转录本中发现了12,000多个m（6）A位点，这些位点具有典型的一致性。位点优先出现在两个不同的标志物终止密码子周围和长的内部外显子内，并且在人类和小鼠之间高度保守。虽然大多数网站都保存良好的正常和癌组织，并在各种刺激，刺激依赖性，动态调制的网站的一个子集被确定。沉默m（6）A甲基转移酶显著影响基因表达和可变剪接模式，导致p53（也称为TP 53）信号通路的调节和细胞凋亡。因此，我们的研究结果表明，RNA修饰的m（6）A在基因表达的调控中发挥着重要作用。
An extensive repertoire of modifications is known to underlie the versatile coding, structural and catalytic functions of RNA, but it remains largely uncharted territory. Although biochemical studies indicate that N-6-methyladenosine (m(6)A) is the most prevalent internal modification in messenger RNA, an in-depth study of its distribution and functions has been impeded by a lack of robust analytical methods. Here we present the human and mouse m(6)A modification landscape in a transcriptome-wide manner, using a novel approach, m(6)A-seq, based on antibody-mediated capture and massively parallel sequencing. We identify over 12,000 m(6)A sites characterized by a typical consensus in the transcripts of more than 7,000 human genes. Sites preferentially appear in two distinct landmarks-around stop codons and within long internal exons-and are highly conserved between human and mouse. Although most sites are well preserved across normal and cancerous tissues and in response to various stimuli, a subset of stimulus-dependent, dynamically modulated sites is identified. Silencing the m(6)A methyltransferase significantly affects gene expression and alternative splicing patterns, resulting in modulation of the p53 (also known as TP53) signalling pathway and apoptosis. Our findings therefore suggest that RNA decoration by m(6)A has a fundamental role in regulation of gene expression.