Transcriptomic profiling of gene expression and RNA processing during Leishmania major differentiation.
Transcriptomic profiling of gene expression and RNA processing during Leishmania major differentiation.
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DOI：
10.1093/nar/gkv656
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发表时间：
2015-08-18
影响因子：
14.9
通讯作者：
El-Sayed NM
中科院分区：
文献类型：
--
作者：
Dillon LA;Okrah K;Hughitt VK;Suresh R;Li Y;Fernandes MC;Belew AT;Corrada Bravo H;Mosser DM;El-Sayed NM
利什曼原虫属的原生动物寄生虫是利什曼病的病原体，利什曼病是一组疾病，全球每年发病率为0.9-1.6百万例。我们使用RNA-seq对基因表达和RNA加工事件的全局变化进行了高分辨率转录组学分析，这些变化发生在L. major从非感染性前环前鞭毛体到感染性后环前鞭毛体的转变中。对多个生物重复进行仔细的统计分析并去除批次效应，为全面分析该病原体获得感染性时的差异基因表达和转录组重塑提供了高质量的框架。我们还确定了大多数利什曼原虫基因的精确5′和3′ UTR边界，并检测到广泛的选择性反式剪接和多聚腺苷酸化。阶段特异性优先反式剪接或多聚腺苷酸化位点和差异表达基因之间可能的相关性的调查显示，缺乏系统的关联，建立表达水平的差异不能归因于阶段调节的替代RNA加工。我们的研究结果建立在现有表达数据集的基础上，并改进了现有的表达数据集，为L. major生物学提供了更详细的信息，这将为该领域提供信息，并可能为药物发现和疫苗开发工作提供更坚实的基础。
Protozoan parasites of the genus Leishmania are the etiological agents of leishmaniasis, a group of diseases with a worldwide incidence of 0.9–1.6 million cases per year. We used RNA-seq to conduct a high-resolution transcriptomic analysis of the global changes in gene expression and RNA processing events that occur as L. major transforms from non-infective procyclic promastigotes to infective metacyclic promastigotes. Careful statistical analysis across multiple biological replicates and the removal of batch effects provided a high quality framework for comprehensively analyzing differential gene expression and transcriptome remodeling in this pathogen as it acquires its infectivity. We also identified precise 5′ and 3′ UTR boundaries for a majority of Leishmania genes and detected widespread alternative trans-splicing and polyadenylation. An investigation of possible correlations between stage-specific preferential trans-splicing or polyadenylation sites and differentially expressed genes revealed a lack of systematic association, establishing that differences in expression levels cannot be attributed to stage-regulated alternative RNA processing. Our findings build on and improve existing expression datasets and provide a substantially more detailed view of L. major biology that will inform the field and potentially provide a stronger basis for drug discovery and vaccine development efforts.