The mRNA-bound proteome of Leishmania mexicana : novel genetic insight into an ancient parasite
The mRNA-bound proteome of Leishmania mexicana : novel genetic insight into an ancient parasite
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墨西哥利什曼原虫的 mRNA 结合蛋白质组：对一种古老寄生虫的新遗传见解
DOI：
10.1101/592402
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发表时间：
2019
期刊：
影响因子：
--
通讯作者：
De Pablos L
中科院分区：
文献类型：
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作者：
De Pablos L
利什曼原虫感染，称为利什曼病，导致全球重大传染病负担。寄生虫的生命周期包含三个主要阶段，需要精确的基因调控协调，才能在白蛉和哺乳动物宿主之间的环境变化中生存。在动质体寄生虫的组成型转录意味着基因调控是压倒性地依赖于转录后机制，但惊人的少数利什曼原虫trans-regulators是已知的。使用优化的交联和深度，定量质谱，我们提出了一个全面的分析1400 mRNA结合蛋白（mRBP）和全细胞蛋白质组从三个主要的利什曼病阶段。支持有效性，尽管交联的RBPome更富集，但交联和非交联的RBPome的蛋白质身份几乎相同。此外，多种候选RBP被内源性标记，并发现以阶段特异性方式与离散mRNA靶池相关联。结果表明，在L.在墨西哥寄生虫中，mRNA水平不是编码蛋白的全细胞表达或RNA结合潜力的强预测因子。证据包括转录本和相应蛋白质表达之间的低相关性以及蛋白质表达versusRNA结合潜力的阶段特异性变化。不出所料，RNA结合蛋白富集与特定生命周期阶段的相对复制效率密切相关。我们的研究是第一个定量定义和比较mRBPome的多个阶段的动质体寄生虫。它提供了新的，深入的见解，trans-regulatory mRNA：蛋白质（mRNP）复合物，驱动利什曼原虫生命周期的进展。
Leishmaniaparasite infections, termed the leishmaniases, cause significant global infectious disease burden. The lifecycle of the parasite embodies three main stages that require precise coordination of gene regulation to survive environmental shifts between sandfly and mammalian hosts. Constitutive transcription in kinetoplastid parasites means that gene regulation is overwhelmingly reliant on post-transcriptional mechanisms, yet strikingly fewLeishmania trans-regulators are known. Using optimized crosslinking and deep, quantified mass spectrometry, we present a comprehensive analysis of 1400 mRNA binding proteins (mRBPs) and whole cell proteomes from the three mainLeishmanialifecycle stages. Supporting the validity, although the crosslinked RBPome is magnitudes more enriched, the protein identities of the crosslinked and non-crosslinked RBPomes were nearly identical. Moreover, multiple candidate RBPs were endogenously tagged and found to associate with discrete mRNA target pools in a stage-specific manner. Results indicate that inL. mexicanaparasites, mRNA levels are not a strong predictor of the whole cell expression or RNA binding potential of encoded proteins. Evidence includes a low correlation between transcript and corresponding protein expression and stage-specific variation in protein expressionversusRNA binding potential. Unsurprisingly, RNA binding protein enrichment correlates strongly with relative replication efficiency of the specific lifecycle stage. Our study is the first to quantitatively define and compare the mRBPome of multiple stages in kinetoplastid parasites. It provides novel, in-depth insight into thetrans-regulatory mRNA:Protein (mRNP) complexes that driveLeishmaniaparasite lifecycle progression.