Simplified RNA secondary structure mapping by automation of SHAPE data analysis.

Simplified RNA secondary structure mapping by automation of SHAPE data analysis.
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DOI:
10.1093/nar/gkr773
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发表时间:
2011-12
影响因子:
14.9
通讯作者:
Glenn JS
Glenn JS
中科院分区:
生物学2区
文献类型:
--
作者:
Pang PS;Elazar M;Pham EA;Glenn JS

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Shape(引物延伸分析的选择性2‘-羟基酰化)技术已经成为在核苷酸水平上确定RNA二级结构的主要方法之一。使用SHAPE的一个重要瓶颈是所需的复杂且耗时的数据处理。我们提出了一种改进的数据收集方法和一系列算法,并在一个名为FAST的程序中实现了该方法和一系列算法,该程序显著减少了处理时间。我们已经使用这种方法来解析丙型肝炎病毒基因组第一个∼900 NT的二级结构,包括整个核心基因。我们还证明了SHAPE/FAST能够检测小分子抑制物与丙型肝炎病毒内部核糖体进入位点(IRES)的结合。总而言之,FAST允许进行高吞吐量的数据处理,以匹配当前SHAPE可能产生的高吞吐量数据,从而降低了确定感兴趣的RNA结构的障碍。
SHAPE (Selective 2′-hydroxyl acylation analysed by primer extension) technology has emerged as one of the leading methods of determining RNA secondary structure at the nucleotide level. A significant bottleneck in using SHAPE is the complex and time-consuming data processing that is required. We present here a modified data collection method and a series of algorithms, embodied in a program entitled Fast Analysis of SHAPE traces (FAST), which significantly reduces processing time. We have used this method to resolve the secondary structure of the first ∼900 nt of the hepatitis C virus (HCV) genome, including the entire core gene. We have also demonstrated the ability of SHAPE/FAST to detect the binding of a small molecule inhibitor to the HCV internal ribosomal entry site (IRES). In conclusion, FAST allows for high-throughput data processing to match the current high-throughput generation of data possible with SHAPE, reducing the barrier to determining the structure of RNAs of interest.
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