Song Jia L, Wessel Gary M
Department of Molecular Biology, Cell Biology and Biochemistry, Brown University, Providence, Rhode Island 02912, USA.
Dev Dyn. 2007 Nov;236(11):3180-90. doi: 10.1002/dvdy.21353.
RNA-mediated interference (RNAi) is a conserved gene silencing mechanism that involves double-stranded RNA as a signal to trigger the sequence-specific degradation of target mRNA, resulting in posttranscriptional silencing and/or translational repression. Bioinformatic searches in the sea urchin genome database identified homologs of Drosha, DGCR5, Dicer, TRBP, Exportin-5, and Argonautes. Quantitative, real-time polymerase chain reaction indicated that all mRNA accumulate in eggs and in variable levels throughout early development. Whole-mount in situ RNA hybridization showed that all of the important players of the RNAi silencing pathway have abundant mRNA accumulation in oocytes and eggs, but have distinct spatial and temporal expression patterns throughout development. Sequence analysis revealed that each of the four Argonautes examined contain conserved residues important for RNAseH activity within the Piwi domain. This study elucidated that genes involved in the RNAi silencing pathway have dynamic expression and, thus, may have regulatory roles during germ cell development and embryogenesis.
RNA介导的干扰(RNAi)是一种保守的基因沉默机制,它涉及双链RNA作为信号来触发靶mRNA的序列特异性降解,导致转录后沉默和/或翻译抑制。在海胆基因组数据库中的生物信息学搜索鉴定出了Drosha、DGCR5、Dicer、TRBP、Exportin-5和Argonaute的同源物。定量实时聚合酶链反应表明,所有mRNA在卵中积累,并在整个早期发育过程中呈现不同水平。全胚胎原位RNA杂交显示,RNAi沉默途径的所有重要参与者在卵母细胞和卵中都有丰富的mRNA积累,但在整个发育过程中具有不同的时空表达模式。序列分析表明,所检测的四个Argonaute中的每一个在Piwi结构域内都含有对RNA酶H活性重要的保守残基。这项研究阐明,参与RNAi沉默途径的基因具有动态表达,因此可能在生殖细胞发育和胚胎发生过程中具有调控作用。