No miRNA were found in Plasmodium and the ones identified in erythrocytes could not be correlated with infection.
No miRNA were found in Plasmodium and the ones identified in erythrocytes could not be correlated with infection.
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DOI：
10.1186/1475-2875-7-47
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发表时间：
2008-03-10
期刊：
影响因子：
3
通讯作者：
Pan W
中科院分区：
文献类型：
--
作者：
Xue X;Zhang Q;Huang Y;Feng L;Pan W
疟原虫在其复杂的生命周期中的转录调控需要不同的遗传程序的顺序激活和/或抑制。microRNA（miRNAs）是一类高度保守的非编码RNA，通过序列特异性抑制基因表达，在调节多种细胞功能中发挥重要作用。疟原虫中的双链RNA介导的基因沉默（RNAi）和转录后基因沉默（PTGS）的知识促使我们推测miRNA是否也可以在疟原虫感染的红细胞中发挥作用。在132个小RNA序列中，没有发现疟原虫特异性的miRNAs，但人类miRNAs miR-451在疟原虫中高表达，约占所有已鉴定miRNAs的1/3。对miR-451表达和疟疾感染的进一步分析显示，miR-451在恶性疟原虫-iRBC中的积累、恶性疟原虫在红细胞中的生命周期阶段或伯氏疟原虫在小鼠中的生命周期阶段之间没有关联。此外，用miR-451的反义寡核苷酸处理对红细胞期恶性疟原虫的生长没有显著影响。在变性聚丙烯酰胺凝胶中分离来自恶性疟原虫-iRBC的混合阶段的短RNA，并克隆到T载体中以创建cDNA文库。然后对单个克隆进行测序，并通过生物信息学预测进行进一步分析，以发现恶性疟原虫-iRBC中可能的miRNA。通过北方印迹和miR-451的反义寡核苷酸（阿索）分析miR-451表达与寄生虫的关系。这些结果有助于消除恶性疟原虫中miRNA的可能性。恶性疟原虫中miRNA的缺失可能与argonaute/dicer基因的缺失相关。此外，miR-451在疟原虫感染的RBC中的积累与寄生虫感染无关。它的积累可能只是红系分化的残留物，也可能是维持成熟红细胞正常功能的成分。
The transcriptional regulation of Plasmodium during its complex life cycle requires sequential activation and/or repression of different genetic programmes. MicroRNAs (miRNAs) are a highly conserved class of non-coding RNAs that are important in regulating diverse cellular functions by sequence-specific inhibition of gene expression. What is know about double-stranded RNA-mediated gene silencing (RNAi) and posttranscriptional gene silencing (PTGS) in Plasmodium parasites entice us to speculate whether miRNAs can also function in Plasmodium-infected RBCs. Of 132 small RNA sequences, no Plasmodium-specific miRNAs have been found. However, a human miRNA, miR-451, was highly expressed, comprising approximately one third of the total identified miRNAs. Further analysis of miR-451 expression and malaria infection showed no association between the accumulation of miR-451 in Plasmodium falciparum-iRBCs, the life cycle stage of P. falciparum in the erythrocyte, or of P. berghei in mice. Moreover, treatment with an antisense oligonucleotide to miR-451 had no significant effect on the growth of the erythrocytic-stage P. falciparum. Short RNAs from a mixed-stage of P. falciparum-iRBC were separated in a denaturing polyacrylamide gel and cloned into T vectors to create a cDNA library. Individual clones were then sequenced and further analysed by bioinformatics prediction to discover probable miRNAs in P. falciparum-iRBC. The association between miR-451 expression and the parasite were analysed by Northern blotting and antisense oligonucleotide (ASO) of miR-451. These results contribute to eliminate the probability of miRNAs in P. falciparum. The absence of miRNA in P. falciparum could be correlated with absence of argonaute/dicer genes. In addition, the miR-451 accumulation in Plasmodium-infected RBCs is independent of parasite infection. Its accumulation might be only the residual of erythroid differentiation or a component to maintain the normal function of mature RBCs.
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