Systematic identification and analysis of exonic splicing silencers
Systematic identification and analysis of exonic splicing silencers
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DOI：
10.1016/j.cell.2004.11.010
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发表时间：
2004-12-17
期刊：
影响因子：
64.5
通讯作者：
Burge, CB
中科院分区：
文献类型：
--
作者：
Wang, ZF;Rolish, ME;Burge, CB
外显子剪接沉默子（ESSs）是抑制邻近剪接位点使用的顺式调控元件，通常导致选择性剪接（AS）。为了系统地鉴定ESSs，我们开发了一个体内剪接报告系统来筛选随机十核苷酸库。屏幕产生了141个ESS十元，其中133个是独特的。在异源外显子/内含子环境和第二种细胞类型中也证实了这些序列的沉默活性。在独特的ESS十十年中，大多数可以聚在一起产生7个假定的ESS基序，其中一些类似于由hnRNPs H和A1结合的已知基序。使用ExonScan算法探索了ESSs在本构剪接中的潜在作用，该算法基于已知或假定的剪接相关基序模拟剪接。ExonScan和相关的生物信息学分析表明，这些ESS基序在抑制假外显子、剪接位点定义和AS中发挥重要作用。
Exonic splicing silencers (ESSs) are cis-regulatory elements that inhibit the use of adjacent splice sites, often contributing to alternative splicing (AS). To systematically identify ESSs, an in vivo splicing reporter system was developed to screen a library of random decanucleotides. The screen yielded 141 ESS decamers, 133 of which were unique. The silencer activity of over a dozen of these sequences was also confirmed in a heterologous exon/intron context and in a second cell type. Of the unique ESS decamers, most could be clustered into groups to yield seven putative ESS motifs, some resembling known motifs bound by hnRNPs H and A1. Potential roles of ESSs in constitutive splicing were explored using an algorithm, ExonScan, which simulates splicing based on known or putative splicing-related motifs. ExonScan and related bioinformatic analyses suggest that these ESS motifs play important roles in suppression of pseudoexons, in splice site definition, and in AS.