Wang Jinkai, Lu Zhi-xiang, Tokheim Collin J, Miller Sara E, Xing Yi
Department of Microbiology, Immunology, and Molecular Genetics, University of California, Los Angeles.
Department of Internal Medicine, University of Iowa.
Mol Biol Evol. 2015 Feb;32(2):481-94. doi: 10.1093/molbev/msu317. Epub 2014 Nov 14.
Changes in exon-intron structures and splicing patterns represent an important mechanism for the evolution of gene functions and species-specific regulatory networks. Although exon creation is widespread during primate and human evolution and has been studied extensively, much less is known about the scope and potential impact of human-specific exon loss events. Historically, transcriptome data and exon annotations are significantly biased toward humans over nonhuman primates. This ascertainment bias makes it challenging to discover human-specific exon loss events. We carried out a transcriptome-wide search of human-specific exon loss events, by taking advantage of RNA sequencing (RNA-seq) as a powerful and unbiased tool for exon discovery and annotation. Using RNA-seq data of humans, chimpanzees, and other primates, we reconstructed and compared transcript structures across the primate phylogeny. We discovered 33 candidate human-specific exon loss events, among which six exons passed stringent experimental filters for the complete loss of splicing activities in diverse human tissues. These events may result from human-specific deletion of genomic DNA, or small-scale sequence changes that inactivated splicing signals. The impact of human-specific exon loss events is predominantly regulatory. Three of the six events occurred in the 5' untranslated region (5'-UTR) and affected cis-regulatory elements of mRNA translation. In SLC7A6, a gene encoding an amino acid transporter, luciferase reporter assays suggested that both a human-specific exon loss event and an independent human-specific single nucleotide substitution in the 5'-UTR increased mRNA translational efficiency. Our study provides novel insights into the molecular mechanisms and evolutionary consequences of exon loss during human evolution.
外显子-内含子结构和剪接模式的变化是基因功能和物种特异性调控网络进化的重要机制。尽管外显子的产生在灵长类和人类进化过程中广泛存在且已得到广泛研究,但对于人类特有的外显子缺失事件的范围和潜在影响却知之甚少。从历史上看,转录组数据和外显子注释在很大程度上偏向人类而非非人类灵长类动物。这种确定偏差使得发现人类特有的外显子缺失事件具有挑战性。我们利用RNA测序(RNA-seq)作为一种强大且无偏差的外显子发现和注释工具,在全转录组范围内搜索人类特有的外显子缺失事件。利用人类、黑猩猩和其他灵长类动物的RNA-seq数据,我们重建并比较了灵长类系统发育中的转录本结构。我们发现了33个候选的人类特有的外显子缺失事件,其中六个外显子通过了严格的实验筛选,证明在多种人类组织中完全丧失了剪接活性。这些事件可能是由于人类特有的基因组DNA缺失,或者是使剪接信号失活的小规模序列变化导致的。人类特有的外显子缺失事件的影响主要是调控性的。六个事件中有三个发生在5'非翻译区(5'-UTR),并影响了mRNA翻译的顺式调控元件。在编码氨基酸转运蛋白的基因SLC7A6中,荧光素酶报告基因实验表明,人类特有的外显子缺失事件和5'-UTR中一个独立的人类特有的单核苷酸替换都提高了mRNA的翻译效率。我们的研究为人类进化过程中外显子缺失的分子机制和进化后果提供了新的见解。