Screening reveals conserved and nonconserved transcriptional regulatory elements including an E3/E4 allele-dependent APOE coding region enhancer
Screening reveals conserved and nonconserved transcriptional regulatory elements including an E3/E4 allele-dependent APOE coding region enhancer
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DOI：
10.1016/j.ygeno.2008.07.009
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发表时间：
2008-11-01
期刊：
影响因子：
4.4
通讯作者：
Smith, Desmond J.
中科院分区：
文献类型：
--
作者：
Chen, Hsuan Pu;Lin, Andy;Smith, Desmond J.
我们利用鸟枪法克隆入荧光素酶载体，在含有人载脂蛋白(APO)E/C1/C4/C2基因簇的153 kb区域进行了无偏倚的实验研究。观察到了转录效应大小的连续体，这可能解释了生物信息学在识别调控区域方面的有限成功。我们确定了9个具有统计学意义的增强子和5个在肝脏或星形胶质细胞中起作用的沉默因子，其中包括两个先前已知的增强子。这14个元件中只有两个含有保守的非编码序列。在APOE基因的编码序列中，我们发现了与阿尔茨海默病相关的E4等位基因的增强子，但不是E3。导致E4/E3氨基酸替换的单核苷酸多态(SNP)是这些变异的原因，这可能解释了E4的高表达水平。我们的结果表明，哺乳动物转录调控序列的种类比目前所认识的更广泛，其中可能包括编码区SNPs。(C)2008 Elsevier Inc.保留所有权利。
We performed an unbiased experimental search for enhancers and silencers in a 153-kb region containing the human apolipoprotein (APO) E/C1/C4/C2 gene cluster using shotgun cloning into a luciferase vector. A continuum of transcriptional effect sizes was observed, possibly explaining the limited success of bioinformatics in identifying regulatory regions. We identified nine statistically significant enhancers and five silencers functional in either liver or astrocyte cells, including two previously known enhancers. Only two of the fourteen elements contained conserved noncoding sequences. Within the coding sequence of the APOE gene we identified an enhancer for the E4 allele associated with Alzheimer's disease, but not E3. The single nucleotide polymorphism (SNP) causing the E4/E3 amino acid substitution was responsible for these variations, potentially explaining the higher expression levels of E4. Our results suggest a wider variety of mammalian transcriptional regulatory sequences than is currently recognized and that these may include coding region SNPs. (C) 2008 Elsevier Inc. All rights reserved.