Comparative ribosome profiling reveals extensive translational complexity in different Trypanosoma brucei life cycle stages.
Comparative ribosome profiling reveals extensive translational complexity in different Trypanosoma brucei life cycle stages.
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DOI：
10.1093/nar/gkt1386
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发表时间：
2014-04
影响因子：
14.9
通讯作者：
Siegel TN
中科院分区：
文献类型：
--
作者：
Vasquez JJ;Hon CC;Vanselow JT;Schlosser A;Siegel TN
虽然基因表达是一个基本的和严格控制的细胞过程，在多个步骤中进行调节，但每个步骤的确切贡献在任何生物体中都是未知的。在原生动物寄生虫布氏锥虫RNA聚合酶II的转录起始调节的情况下，大大简化了阐明翻译对全球基因表达的贡献的任务。因此，我们对T.布氏杆菌，允许对RNA翻译和翻译效率进行全基因组分析。我们发现，后者之间的寄生虫的生命周期阶段和100倍的基因之间的差异很大，从而有助于基因表达的RNA稳定性的类似程度。在子密码子分辨率下定位核糖体位置的能力揭示了位于5′ UTR内的上游开放阅读框的广泛翻译，并使数百个先前未注释的推定编码序列（CDS）的鉴定成为可能。对现有蛋白质组学和全基因组RNAi数据的评估证实了先前未注释的CDS的翻译，并表明>200个这些CDS在寄生虫存活中的重要作用，特别是以感染哺乳动物的形式。总之，我们的数据表明，翻译控制在T.布鲁塞。
While gene expression is a fundamental and tightly controlled cellular process that is regulated at multiple steps, the exact contribution of each step remains unknown in any organism. The absence of transcription initiation regulation for RNA polymerase II in the protozoan parasite Trypanosoma brucei greatly simplifies the task of elucidating the contribution of translation to global gene expression. Therefore, we have sequenced ribosome-protected mRNA fragments in T. brucei, permitting the genome-wide analysis of RNA translation and translational efficiency. We find that the latter varies greatly between life cycle stages of the parasite and ∼100-fold between genes, thus contributing to gene expression to a similar extent as RNA stability. The ability to map ribosome positions at sub-codon resolution revealed extensive translation from upstream open reading frames located within 5′ UTRs and enabled the identification of hundreds of previously un-annotated putative coding sequences (CDSs). Evaluation of existing proteomics and genome-wide RNAi data confirmed the translation of previously un-annotated CDSs and suggested an important role for >200 of those CDSs in parasite survival, especially in the form that is infective to mammals. Overall our data show that translational control plays a prevalent and important role in different parasite life cycle stages of T. brucei.
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