RNA motif discovery by SHAPE and mutational profiling (SHAPE-MaP).

RNA motif discovery by SHAPE and mutational profiling (SHAPE-MaP).
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DOI:
10.1038/nmeth.3029
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发表时间:
2014-09
期刊:
影响因子:
48
通讯作者:
Weeks, Kevin M.
Weeks, Kevin M.
中科院分区:
生物学1区
文献类型:
--
作者:
Siegfried, Nathan A.;Busan, Steven;Rice, Greggory M.;Nelson, Julie A. E.;Weeks, Kevin M.

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许多生物过程是RNA介导的,但大多数RNA的高阶结构是未知的,这使得很难理解RNA结构如何控制功能。在这里,我们描述了SHAPE突变谱(SHAPE- map),使RNA功能基序的重新和大规模鉴定成为可能。通过大规模平行测序测量,在cDNA合成过程中,SHAPE的2 ' -羟基酰化位点被编码为非互补核苷酸。shape - map引导的建模识别了已知结构复杂RNA中90%以上的可接受碱基对,并用于定义HIV-1 RNA基因组的第二代模型。HIV-1模型包含所有已知的结构基序和以前未知的元素,包括实验验证的假结。SHAPE-MaP产生准确和高分辨率的二级结构模型,能够分析低丰度RNA,在单次实验中解开序列多态性,并最终使RNA结构分析民主化。
Many biological processes are RNA-mediated, but higher-order structures for most RNAs are unknown, making it difficult to understand how RNA structure governs function. Here we describe SHAPE mutational profiling (SHAPE-MaP) that makes possible de novo and large-scale identification of RNA functional motifs. Sites of 2’-hydroxyl acylation by SHAPE are encoded as non-complementary nucleotides during cDNA synthesis, as measured by massively parallel sequencing. SHAPE-MaP-guided modeling identified greater than 90% of accepted base pairs in complex RNAs of known structure and was used to define a second-generation model for the HIV-1 RNA genome. The HIV-1 model contains all known structured motifs and previously unknown elements, including experimentally validated pseudoknots. SHAPE-MaP yields accurate and high-resolution secondary structure models, enables analysis of low abundance RNAs, disentangles sequence polymorphisms in single experiments, and will ultimately democratize RNA structure analysis.
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发表时间: 2014-06
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影响因子: --
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