Sanders Matthew, Maddelein Wendy, Depicker Anna, Van Montagu Marc, Cornelissen Marc, Jacobs John
Department of Plant Systems Biology, Flanders Interuniversity Institute for Biotechnology (VIB), Ghent University, K.L. Ledeganckstraat 35, Belgium.
EMBO J. 2002 Nov 1;21(21):5824-32. doi: 10.1093/emboj/cdf586.
Post-transcriptional gene silencing (PTGS) is characterized by the accumulation of short interfering RNAs that are proposed to mediate sequence-specific degradation of cognate and secondary target mRNAs. In plants, it is unclear to what extent endogenous genes contribute to this process. Here, we address the role of the endogenous target genes in transgene-mediated PTGS of beta-1,3-glucanases in tobacco. We found that mRNA sequences of the endogenous glucanase glb gene with varying degrees of homology to the Nicotiana plumbaginifolia gn1 transgene are targeted by the silencing machinery, although less efficiently than corresponding transgene regions. Importantly, we show that endogene-specific nucleotides in the glb sequence provide specificity to the silencing process. Consistent with this finding, small sense and antisense 21- to 23-nucleotide RNAs homologous to the endogenous glb gene were detected. Combined, these data demonstrate that a co-suppressed endogenous glucan ase gene is involved in signal amplification and selection of homologous targets, and show that endogenous genes can actively participate in PTGS in plants. The findings are introduced as a further sophistication of the post-transciptional silencing model.
转录后基因沉默(PTGS)的特征是短干扰RNA的积累,这些短干扰RNA被认为介导同源和次级靶标mRNA的序列特异性降解。在植物中,内源性基因在这一过程中所起的作用尚不清楚。在此,我们探讨了内源性靶基因在烟草中β-1,3-葡聚糖酶转基因介导的PTGS中的作用。我们发现,与烟草垂花烟草gn1转基因具有不同程度同源性的内源性葡聚糖酶glb基因的mRNA序列被沉默机制靶向,尽管效率低于相应的转基因区域。重要的是,我们表明glb序列中的内源性基因特异性核苷酸为沉默过程提供了特异性。与此发现一致,检测到与内源性glb基因同源的小正义和反义21至23个核苷酸的RNA。综合这些数据表明,一个共抑制的内源性葡聚糖酶基因参与信号放大和同源靶标的选择,并表明内源性基因可以在植物中积极参与PTGS。这些发现被作为转录后沉默模型的进一步完善而引入。