Department of Genome Organization, Engelhardt Institute of Molecular Biology, Russian Academy of Sciences, Moscow, Russia.
PLoS One. 2011;6(7):e21882. doi: 10.1371/journal.pone.0021882. Epub 2011 Jul 14.
In the Drosophila melanogaster germline, the piRNA pathway silences retrotransposons as well as other transcribed repetitive elements. Suffix is an unusual short retroelement that was identified both as an actively transcribed repetitive element and also as an element at the 3' ends of the Drosophila non-LTR F element. The copies of suffix that are F element-independent are far more actively transcribed than their counterparts on the F element. We studied the patterns of small RNAs targeting both strands of suffix in Drosophila ovaries using an RNase protection assay and the analysis of the corresponding RNA sequences from the libraries of total small RNAs. Our results indicate that suffix sense and antisense transcripts are targeted mainly by 23-29 nucleotides in length piRNAs and also by 21 nucleotides in length siRNAs. Suffix sense transcripts actively form longer RNA species, corresponding either to partial digestion products of the RNAi and Piwi pathways or to another RNA silencing mechanism. Both sense and antisense suffix transcripts accumulated in the ovaries of homozygous spn-E, piwi and aub mutants. These results provide evidence that suffix sense and antisense transcripts in the germ line and soma are targeted by both RNAi and Piwi pathways and that a Dicer-independent pathway of biogenesis of siRNAs could exist in Drosophila cells.
在黑腹果蝇生殖系中,piRNA 途径沉默反转录转座子和其他转录重复元件。后缀是一种不寻常的短反转元件,它既是一种活跃转录的重复元件,也是果蝇非 LTR F 因子 3' 末端的一个元件。与 F 因子上的拷贝相比,独立于后缀的拷贝转录更为活跃。我们使用 RNase 保护分析和从小 RNA 总文库中分析相应的 RNA 序列,研究了果蝇卵巢中靶向后缀两条链的小 RNA 的模式。我们的结果表明,后缀的 sense 和 antisense 转录物主要被 23-29 个核苷酸长的 piRNAs 和 21 个核苷酸长的 siRNAs 靶向。后缀 sense 转录物积极地形成更长的 RNA 种类,这些 RNA 种类对应于 RNAi 和 Piwi 途径的部分消化产物或另一种 RNA 沉默机制。后缀 sense 和 antisense 转录物在 spn-E、piwi 和 aub 突变体的卵巢中积累。这些结果表明,生殖系和体中的后缀 sense 和 antisense 转录物都被 RNAi 和 Piwi 途径靶向,并且在果蝇细胞中可能存在一种 Dicer 非依赖性 siRNA 生物发生途径。