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Arabidopsis HEN1: A genetic link between endogenous miRNA controlling development and siRNA controlling transgene silencing and virus resistance

机译:拟南芥HEN1:控制发育的内源性miRNA与控制转基因沉默和病毒抗性的siRNA之间的遗传联系

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摘要

In animals, double-stranded short interfering RNA (siRNA) and single-stranded microRNA (miRNA) regulate gene expression by targeting homologous mRNA for cleavage or by interfering with their translation, respectively [1-3]. siRNAs are processed from injected or transgene-derived, long, perfect double-stranded RNA (dsRNA), while miRNAs are processed from short, imperfect dsRNA precursors transcribed from endogenous intergenic regions [4-9]. In plants, both siRNAs and miRNAs activate cleavage of homologous RNA targets [10-12], but little is known about the genes controlling their production or action. The SGS2/SDE1 protein contributes to produce transgene siRNA [10], while DCL1 and HEN1 contribute to endogenous miRNA accumulation [8, 9]. Here, we show that: I) SGS2, SGS3 [13], AGO1 [14,15], and HEN1 contribute to produce transgene siRNA involved in sense post-transcriptional gene silencing (S-PTGS); ii) HEN1, but not SGS2, SGS3, or AGO1, contributes to the accumulation of the endogenous miR171 miRNA and to the cleavage of Scarecrow target mRNA by miR171 [11]; iii) SGS2, SGS3, AGO1, and HEN I contribute to resistance against cucumber mosaic virus [13, 16], but not to siRNA and IR-PTGS triggered by hairpin transgenes directly producing perfect dsRNA [16]; and iv) the actions of HEN1 in MiRNA/development and siRNA/ S-PTGS can be uncoupled by single-point mutations at different positions in the protein. [References: 30]
机译:在动物中,双链短干扰RNA(siRNA)和单链微RNA(miRNA)分别通过靶向同源mRNA进行切割或干扰其翻译来调节基因表达[1-3]。 siRNA是从注射的或转基因的,长的,完美的双链RNA(dsRNA)中加工而来的,而miRNA是从从内源性基因间区域转录的短而不完美的dsRNA前体来加工的[4-9]。在植物中,siRNA和miRNA均能激活同源RNA靶标的切割[10-12],但对于控制其产生或作用的基因知之甚少。 SGS2 / SDE1蛋白有助于产生转基因siRNA [10],而DCL1和HEN1有助于内源性miRNA积累[8,9]。在这里,我们显示:I)SGS2,SGS3 [13],AGO1 [14,15]和HEN1有助于产生涉及有义转录后基因沉默(S-PTGS)的转基因siRNA; ii)HEN1,而非SGS2,SGS3或AGO1,有助于内源性miR171 miRNA的积累和miR171对稻草人靶mRNA的切割[11]; iii)SGS2,SGS3,AGO1和HEN I对黄瓜花叶病毒有抗药性[13,16],但对直接产生完美dsRNA的发夹转基因触发的siRNA和IR-PTGS无效[16]; iv)HEN1在MiRNA /发育和siRNA / S-PTGS中的作用可以通过蛋白质中不同位置的单点突变来消除。 [参考:30]

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