Wang Xinye, Xu Xindong, Lu Xingyu, Zhang Yuanbin, Pan Weiqing
Institute for Infectious Diseases and Vaccine Development, Tongji University School of Medicine, Shanghai, China.
School of Life Science and Biotechnology, Shanghai Jiao Tong University, Shanghai, China.
PLoS One. 2015 Sep 25;10(9):e0138470. doi: 10.1371/journal.pone.0138470. eCollection 2015.
Alternative splicing is a molecular process that contributes greatly to the diversification of proteome and to gene functions. Understanding the mechanisms of stage-specific alternative splicing can provide a better understanding of the development of eukaryotes and the functions of different genes. Schistosoma japonicum is an infectious blood-dwelling trematode with a complex lifecycle that causes the tropical disease schistosomiasis. In this study, we analyzed the transcriptome of Schistosoma japonicum to discover alternative splicing events in this parasite, by applying RNA-seq to cDNA library of adults and schistosomula. Results were validated by RT-PCR and sequencing. We found 11,623 alternative splicing events among 7,099 protein encoding genes and average proportion of alternative splicing events per gene was 42.14%. We showed that exon skip is the most common type of alternative splicing events as found in high eukaryotes, whereas intron retention is the least common alternative splicing type. According to intron boundary analysis, the parasite possesses same intron boundaries as other organisms, namely the classic "GT-AG" rule. And in alternative spliced introns or exons, this rule is less strict. And we have attempted to detect alternative splicing events in genes encoding proteins with signal peptides and transmembrane helices, suggesting that alternative splicing could change subcellular locations of specific gene products. Our results indicate that alternative splicing is prevalent in this parasitic worm, and that the worm is close to its hosts. The revealed secretome involved in alternative splicing implies new perspective into understanding interaction between the parasite and its host.
可变剪接是一种分子过程,对蛋白质组的多样化和基因功能有很大贡献。了解阶段特异性可变剪接的机制有助于更好地理解真核生物的发育以及不同基因的功能。日本血吸虫是一种寄生在血液中的传染性吸虫,具有复杂的生命周期,可导致热带疾病血吸虫病。在本研究中,我们通过对成虫和童虫的cDNA文库进行RNA测序,分析了日本血吸虫的转录组,以发现该寄生虫中的可变剪接事件。结果通过RT-PCR和测序进行验证。我们在7099个蛋白质编码基因中发现了11623个可变剪接事件,每个基因的可变剪接事件平均比例为42.14%。我们发现,外显子跳跃是高等真核生物中最常见的可变剪接事件类型,而内含子保留是最不常见的可变剪接类型。根据内含子边界分析,该寄生虫具有与其他生物相同的内含子边界,即经典的“GT-AG”规则。在可变剪接的内含子或外显子中,该规则不太严格。我们还试图检测编码具有信号肽和跨膜螺旋的蛋白质的基因中的可变剪接事件,这表明可变剪接可能会改变特定基因产物的亚细胞定位。我们的结果表明,可变剪接在这种寄生蠕虫中很普遍,并且该蠕虫与宿主关系密切。可变剪接所涉及的分泌蛋白组揭示了理解寄生虫与其宿主之间相互作用的新视角。