Characterization of the RNA-Binding Protein TcSgn1 in Trypanosoma cruzi.
Characterization of the RNA-Binding Protein TcSgn1 in Trypanosoma cruzi.
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DOI：
10.3390/microorganisms9050986
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发表时间：
2021-05-02
期刊：
影响因子：
4.5
通讯作者：
Alves LR
中科院分区：
文献类型：
--
作者：
Oliveira C;Gerber AP;Goldenberg S;Alves LR
RNA 结合蛋白 (RBP) 参与基因表达转录后调控的多个步骤，例如剪接、信使 RNA 运输、mRNA 定位和翻译。锥虫的基因表达调控主要发生在转录后水平，RBP 在此过程中发挥着重要作用。在这里，我们表征了 RBP TcSgn1，它包含一个 RNA 识别基序 (RRM)。 TcSgn1 是酵母酿酒酵母蛋白 ScSgn1 的密切直系同源物，在细胞质中的翻译调节中发挥作用。我们发现克氏锥虫中的 TcSgn1 位于指数生长的上鞭毛体的细胞核中。通过对 TcSgn1 进行免疫沉淀测定，我们鉴定了数百个与该蛋白质相关的 mRNA，其中很大一部分编码核酸结合、转录和内吞作用蛋白质。此外，我们还发现 TcSgn1 能够直接与 mRNA 的 Poly(A) 尾相互作用。对营养胁迫下寄生虫的研究表明，TcSgn1 除了定位于细胞核外，还定位于细胞质颗粒中。与ScSgn1类似，我们观察到TcSgn1也与PABP1蛋白相互作用，表明该蛋白可能在调节T. cruzi的基因表达中发挥作用。总而言之，我们的结果表明 RNA 结合蛋白 TcSgn1 是与核功能、应激反应和 RNA 代谢相关的核糖核蛋白复合物的一部分。
RNA-binding proteins (RBPs) participate in several steps of post-transcriptional regulation of gene expression, such as splicing, messenger RNA transport, mRNA localization, and translation. Gene-expression regulation in trypanosomatids occurs primarily at the post-transcriptional level, and RBPs play important roles in the process. Here, we characterized the RBP TcSgn1, which contains one RNA recognition motif (RRM). TcSgn1 is a close ortholog of yeast Saccharomyces cerevisiae protein ScSgn1, which plays a role in translational regulation in the cytoplasm. We found that TcSgn1 in Trypanosoma cruzi is localized in the nucleus in exponentially growing epimastigotes. By performing immunoprecipitation assays of TcSgn1, we identified hundreds of mRNAs associated with the protein, a significant fraction of them coding for nucleic acids binding, transcription, and endocytosis proteins. In addition, we show that TcSgn1 is capable of interacting directly with the poly(A) tail of the mRNAs. The study of parasites under nutritional stress showed that TcSgn1 was localized in cytoplasmic granules in addition to localizing in the nucleus. Similar to ScSgn1, we observed that TcSgn1 also interacts with the PABP1 protein, suggesting that this protein may play a role in regulating gene expression in T. cruzi. Taken together, our results show that RNA-binding protein TcSgn1 is part of ribonucleoprotein complexes associated with nuclear functions, stress response, and RNA metabolism.
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