Division of Biological Sciences, University of Missouri, Columbia, Missouri 65211, USA.
Genetics. 2011 Aug;188(4):823-34. doi: 10.1534/genetics.111.128470. Epub 2011 May 19.
RNA interference (RNAi) regulates gene expression by sequence-specific destruction of RNA. It acts as a defense mechanism against viruses and represses the expression of transposable elements (TEs) and some endogenous genes. We report that mutations and transgene constructs that condition cell death suppress RNA interference in adjacent cells in Drosophila melanogaster. The reversal of RNAi is effective for both the white (w) eye color gene and green fluorescent protein (GFP), indicating the generality of the inhibition. Antiapoptotic transgenes that reverse cell death will also reverse the inhibition of RNAi. Using GFP and a low level of cell death produced by a heat shock-head involution defective (hs-hid) transgene, the inhibition appears to occur by blocking the conversion of double-stranded RNA (dsRNA) to short interfering RNA (siRNA). We also demonstrate that the mus308 gene and endogenous transposable elements, which are both regularly silenced by RNAi, are increased in expression and accompanied by a reduced level of siRNA, when cell death occurs. The finding that chronic ectopic cell death affects RNAi is critical for an understanding of the application of the technique in basic and applied studies. These results also suggest that developmental perturbations, disease states, or environmental insults that cause ectopic cell death would alter transposon and gene expression patterns in the organism by the inhibition of small RNA silencing processes.
RNA 干扰 (RNAi) 通过 RNA 的序列特异性降解来调节基因表达。它作为一种防御机制,可对抗病毒,并抑制转座元件 (TEs) 和一些内源性基因的表达。我们报告称,在果蝇中,可导致细胞死亡的突变和转基因构建物会抑制邻近细胞中的 RNA 干扰。RNAi 的逆转对白色 (w) 眼色基因和绿色荧光蛋白 (GFP) 都有效,表明这种抑制作用具有普遍性。逆转细胞死亡的抗凋亡转基因也会逆转 RNAi 的抑制作用。使用 GFP 和由热休克-头凹陷缺陷 (hs-hid) 转基因产生的低水平细胞死亡,抑制作用似乎是通过阻止双链 RNA (dsRNA) 转化为小干扰 RNA (siRNA) 来实现的。我们还证明,当发生细胞死亡时,mus308 基因和内源性转座元件的表达增加,同时 siRNA 水平降低。细胞死亡会持续影响 RNAi 的发现,对于理解该技术在基础和应用研究中的应用至关重要。这些结果还表明,发育异常、疾病状态或导致异位细胞死亡的环境损伤会通过抑制小 RNA 沉默过程来改变生物体中转座子和基因表达模式。