Li Junjie, Yang Zhiyong, Yu Bin, Liu Jun, Chen Xuemei
Department of Botany and Plant Sciences, Institute of Integrative Genome Research, University of California, Riverside, Riverside, California 92508, USA.
Curr Biol. 2005 Aug 23;15(16):1501-7. doi: 10.1016/j.cub.2005.07.029.
Small RNAs of 21-25 nucleotides (nt), including small interfering RNAs (siRNAs) and microRNAs (miRNAs), act as guide RNAs to silence target-gene expression in a sequence-specific manner. In addition to a Dicer homolog, DCL1, the biogenesis of miRNAs in Arabidopsis requires another protein, HEN1. miRNAs are reduced in abundance and increased in size in hen1 mutants. We found that HEN1 is a miRNA methyltransferase that adds a methyl group to the 3'-most nucleotide of miRNAs, but the role of miRNA methylation was unknown. Here, we show that siRNAs from sense transgenes, hairpin transgenes, and transposons or repeat sequences, as well as a new class of siRNAs known as trans-acting siRNAs, are also methylated in vivo by HEN1. In addition, we show that the size increase of small RNAs in the hen1-1 mutant is due to the addition of one to five U residues to the 3' ends of the small RNAs. Therefore, a novel uridylation activity targets the 3' ends of unmethylated miRNAs and siRNAs in hen1 mutants. We conclude that 3'-end methylation is a common step in miRNA and siRNA metabolism and likely protects the 3' ends of the small RNAs from the uridylation activity.
21至25个核苷酸(nt)的小RNA,包括小干扰RNA(siRNA)和微小RNA(miRNA),作为引导RNA以序列特异性方式使靶基因表达沉默。除了Dicer同源物DCL1外,拟南芥中miRNA的生物合成还需要另一种蛋白质HEN1。在hen1突变体中,miRNA的丰度降低且大小增加。我们发现HEN1是一种miRNA甲基转移酶,可将甲基添加到miRNA的最末端3'核苷酸上,但miRNA甲基化的作用尚不清楚。在这里,我们表明来自正义转基因、发夹转基因、转座子或重复序列的siRNA,以及一类新的称为反式作用siRNA的siRNA,在体内也被HEN1甲基化。此外,我们表明hen1-1突变体中小RNA的大小增加是由于在小RNA的3'末端添加了1至5个U残基。因此,一种新的尿苷化活性靶向hen1突变体中未甲基化的miRNA和siRNA的3'末端。我们得出结论,3'末端甲基化是miRNA和siRNA代谢中的一个常见步骤,可能保护小RNA的3'末端免受尿苷化活性的影响。