Kreuze Jan F, Savenkov Eugene I, Cuellar Wilmer, Li Xiangdong, Valkonen Jari P T
Department of Plant Biology and Forest Genetics, SLU, Uppsala, Sweden.
J Virol. 2005 Jun;79(11):7227-38. doi: 10.1128/JVI.79.11.7227-7238.2005.
Double-stranded RNA (dsRNA)-specific endonucleases belonging to RNase III classes 3 and 2 process dsRNA precursors to small interfering RNA (siRNA) or microRNA, respectively, thereby initiating and amplifying RNA silencing-based antiviral defense and gene regulation in eukaryotic cells. However, we now provide evidence that a class 1 RNase III is involved in suppression of RNA silencing. The single-stranded RNA genome of sweet potato chlorotic stunt virus (SPCSV) encodes an RNase III (RNase3) homologous to putative class 1 RNase IIIs of unknown function in rice and Arabidopsis. We show that RNase3 has dsRNA-specific endonuclease activity that enhances the RNA-silencing suppression activity of another protein (p22) encoded by SPCSV. RNase3 and p22 coexpression reduced siRNA accumulation more efficiently than p22 alone in Nicotiana benthamiana leaves expressing a strong silencing inducer (i.e., dsRNA). RNase3 did not cause intracellular silencing suppression or reduce accumulation of siRNA in the absence of p22 or enhance silencing suppression activity of a protein encoded by a heterologous virus. No other known RNA virus encodes an RNase III or uses two independent proteins cooperatively for RNA silencing suppression.
属于第3类和第2类核糖核酸酶III的双链RNA(dsRNA)特异性核酸内切酶分别将dsRNA前体加工成小干扰RNA(siRNA)或微小RNA,从而在真核细胞中启动并放大基于RNA沉默的抗病毒防御和基因调控。然而,我们现在提供证据表明第1类核糖核酸酶III参与了对RNA沉默的抑制。甘薯褪绿矮缩病毒(SPCSV)的单链RNA基因组编码一种核糖核酸酶III(核糖核酸酶3),与水稻和拟南芥中功能未知的假定第1类核糖核酸酶III同源。我们表明,核糖核酸酶3具有dsRNA特异性核酸内切酶活性,可增强SPCSV编码的另一种蛋白质(p22)的RNA沉默抑制活性。在表达强沉默诱导剂(即dsRNA)的本氏烟草叶片中,核糖核酸酶3和p22共表达比单独表达p22更有效地减少了siRNA的积累。在没有p22的情况下,核糖核酸酶3不会引起细胞内沉默抑制或减少siRNA的积累,也不会增强异源病毒编码的蛋白质的沉默抑制活性。没有其他已知的RNA病毒编码核糖核酸酶III或协同使用两种独立的蛋白质进行RNA沉默抑制。