Monsanto, Calgene Campus, Davis, CA 95616, USA.
Proc Natl Acad Sci U S A. 2011 Jan 4;108(1):409-14. doi: 10.1073/pnas.1009805108. Epub 2010 Dec 20.
In plants, small interfering RNAs (siRNAs) with sequence homology to transcribed regions of genes can guide the sequence-specific degradation of corresponding mRNAs, leading to posttranscriptional gene silencing (PTGS). The current consensus is that siRNA-mediated PTGS occurs primarily in the cytoplasm where target mRNAs are localized and translated into proteins. However, expression of an inverted-repeat double-stranded RNA corresponding to the soybean FAD2-1A desaturase intron is sufficient to silence FAD2-1, implicating nuclear precursor mRNA (pre-mRNA) rather than cytosolic mRNA as the target of PTGS. Silencing FAD2-1 using intronic or 3'-UTR sequences does not affect transcription rates of the target genes but results in the strong reduction of target transcript levels in the nucleus. Moreover, siRNAs corresponding to pre-mRNA-specific sequences accumulate in the nucleus. In Arabidopsis, we find that two enzymes involved in PTGS, Dicer-like 4 and RNA-dependent RNA polymerase 6, are localized in the nucleus. Collectively, these results demonstrate that siRNA-directed RNA degradation can take place in the nucleus, suggesting the need for a more complex view of the subcellular compartmentation of PTGS in plants.
在植物中,与基因转录区域具有序列同源性的小干扰 RNA(siRNA)可以引导相应的 mRNA 进行序列特异性降解,导致转录后基因沉默(PTGS)。目前的共识是,siRNA 介导的 PTGS 主要发生在细胞质中,靶 mRNA 位于细胞质中并翻译成蛋白质。然而,表达与大豆 FAD2-1A 去饱和酶内含子对应的反向重复双链 RNA 足以沉默 FAD2-1,暗示核前体 mRNA(pre-mRNA)而不是细胞质 mRNA 是 PTGS 的靶标。使用内含子或 3'-UTR 序列沉默 FAD2-1 不会影响靶基因的转录率,但会导致靶转录本水平在核内强烈降低。此外,与 pre-mRNA 特异性序列对应的 siRNA 积累在核内。在拟南芥中,我们发现参与 PTGS 的两种酶,Dicer-like 4 和 RNA 依赖性 RNA 聚合酶 6,定位于核内。综上所述,这些结果表明 siRNA 指导的 RNA 降解可以在核内发生,这表明需要对植物中 PTGS 的亚细胞区室化有更复杂的认识。