Molecular characterization of the Trypanosoma cruzi specific RNA binding protein TcRBP40 and its associated mRNAs
Molecular characterization of the Trypanosoma cruzi specific RNA binding protein TcRBP40 and its associated mRNAs
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DOI：
10.1016/j.bbrc.2012.02.154
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发表时间：
2012-04-06
影响因子：
3.1
通讯作者：
Dallagiovanna, Bruno
中科院分区：
文献类型：
--
作者：
Guerra-Slompo, Eloise P.;Probst, Christian M.;Dallagiovanna, Bruno
克氏锥虫是恰加斯病的病原体，这是一种被忽视的疾病，影响着美洲数百万人。克氏锥虫主要依靠转录后调控来控制阶段特异性基因表达。 RNA 结合蛋白 (RBP) 与功能相关的 mRNA 结合，形成定义转录后操纵子的核糖核蛋白复合物。 RNA 识别基序 (RRM) 是最常见、最古老的 RBP 家族。该 RBP 家族已在锥虫寄生虫中得到鉴定，但仅对其中少数进行了功能表征。我们在此描述了 TcRBP40（克氏锥虫特异性 RBP）及其相关 mRNA 的功能特征。我们使用重组 RIP-Chip 测定的修改版本来鉴定与其关联的 mRNA，并使用体内 TAP 标签测定来确认这些结果。 TcRBP40 与相关 mRNA 3'UTR 中富含 AG 的序列结合，发现这些序列主要编码假定的跨膜蛋白。 TcRBP40 在后生发育过程中差异表达。令人惊讶的是，在上鞭毛体中，它分散在细胞质中，但集中在储库体（一种克氏锥虫特异性细胞器）中，这表明这种寄生虫细胞器具有假定的新功能。 (C) 2012 Elsevier Inc. 保留所有权利。
Trypanosoma cruzi is the causative agent of Chagas disease, a neglected disorder that affects millions of people in the Americas. T. cruzi relies mostly upon post-transcriptional regulation to control stage specific gene expression. RNA binding proteins (RBPs) associate with functionally related mRNAs forming ribonucleoprotein complexes that define post-transcriptional operons. The RNA Recognition Motif (RRM) is the most common and ancient family of RBPs. This family of RBPs has been identified in trypanosomatid parasites and only a few of them have been functionally characterized. We describe here the functional characterization of TcRBP40, a T. cruzi specific RBP, and its associated mRNAs. We used a modified version of the recombinant RIP-Chip assay to identify the mRNAs with which it associates and in vivo TAP-tag assays to confirm these results. TcRBP40 binds to an AG-rich sequence in the 3'UTR of the associated mRNAs, which were found to encode mainly putative transmembrane proteins. TcRBP40 is differentially expressed in metacyclogenesis. Surprisingly, in epimastigotes, it is dispersed in the cytoplasm but is concentrated in the reservosomes, a T. cruzi specific organelle, which suggests a putative new function for this parasite organelle. (C) 2012 Elsevier Inc. All rights reserved.