Costa Jimena Ferreira da, Ferrarini Mariana Galvão, Nardelli Sheila Cristina, Goldenberg Samuel, Ávila Andréa Rodrigues, Holetz Fabíola Barbieri
Instituto Carlos Chagas, Fundação Oswaldo Cruz-Fiocruz, Curitiba, PR, Brasil.
Mem Inst Oswaldo Cruz. 2018;113(6):e170531. doi: 10.1590/0074-02760170531. Epub 2018 Jun 18.
BACKGROUND Eukaryotic ribonucleoprotein (RNP) granules are important for the regulation of RNA fate. RNP granules exist in trypanosomatids; however, their roles in controlling gene expression are still not understood. XRNA is a component of granules in Trypanosoma brucei but has not been investigated in Trypanosoma cruzi. OBJECTIVES This study aimed to investigate the TcXRNA dynamic assembly and its interaction with RNP components under conditions that affect the mRNA availability. METHODS We used in vitro metacyclogenesis of T. cruzi to observe changes in RNP granules during the differentiation process. TcXRNA expression was analysed by Western blot and immunofluorescence. Colocalisation assays were performed to investigate the interaction of TcXRNA with other RNP components. FINDINGS TcXRNA is constantly present during metacyclogenesis and is localised in cytoplasmic granules. TcXRNA does not colocalise with TcDHH1 and TcCAF1 granules in the cytoplasm. However, TcXRNA granules colocalise with mRNP granules at the nuclear periphery when mRNA processing is inhibited. MAIN CONCLUSIONS TcXRNA plays a role in mRNA metabolism as a component of mRNP granules whose assembly is dependent on mRNA availability. TcXRNA granules colocalise with distinct RNP granules at the nuclear periphery, suggesting that the perinuclear region is a regulatory compartment in T. cruzi mRNA metabolism.
背景 真核核糖核蛋白(RNP)颗粒对RNA命运的调控至关重要。RNP颗粒存在于锥虫中;然而,它们在控制基因表达中的作用仍不清楚。XRNA是布氏锥虫颗粒的一个组成部分,但尚未在克氏锥虫中进行研究。
目的 本研究旨在研究克氏锥虫TcXRNA的动态组装及其在影响mRNA可用性的条件下与RNP成分的相互作用。
方法 我们利用克氏锥虫的体外循环后期分化来观察分化过程中RNP颗粒的变化。通过蛋白质免疫印迹和免疫荧光分析TcXRNA的表达。进行共定位分析以研究TcXRNA与其他RNP成分的相互作用。
研究结果 TcXRNA在循环后期分化过程中持续存在,并定位于细胞质颗粒中。TcXRNA在细胞质中不与TcDHH1和TcCAF1颗粒共定位。然而,当mRNA加工受到抑制时,TcXRNA颗粒在核周与mRNP颗粒共定位。
主要结论 TcXRNA作为mRNP颗粒的一个组成部分在mRNA代谢中发挥作用,其组装依赖于mRNA的可用性。TcXRNA颗粒在核周与不同的RNP颗粒共定位,表明核周区域是克氏锥虫mRNA代谢中的一个调控区室。