Zrachya Avi, Glick Efrat, Levy Yael, Arazi Tzahi, Citovsky Vitaly, Gafni Yedidya
Institute of Plant Sciences, Agricultural Research Organization, Volcani Center, P.O. Box 6, Bet Dagan 50250, Israel.
Virology. 2007 Feb 5;358(1):159-65. doi: 10.1016/j.virol.2006.08.016. Epub 2006 Sep 18.
The Israeli isolate of Tomato yellow leaf curl geminivirus (TYLCV-Is) is a major tomato pathogen, causing extensive crop losses both in the New and Old World. Surprisingly, however, little is known about the molecular mechanisms of TYLCV-Is interactions with tomato cells. Here, we have identified a TYLCV-Is protein, V2, which acts as a suppressor of RNA silencing and which is unrelated to presently known viral suppressors. Specifically, V2, but not other proteins of TYLCV-Is, inhibited RNA silencing of a reporter transgene, GFP. This inhibition elevated the cellular levels of the GFP transcript and the GFP protein, but it had no apparent effect on the accumulation of GFP-specific short interfering RNAs (siRNAs), suggesting that TYLCV-Is V2 targets a step in the RNA silencing pathway which is subsequent to the Dicer-mediated cleavage of dsRNA. Visualization of the sub-cellular localization of TYLCV-Is V2 in plant protoplasts and tissues showed that this protein is associated with cytoplasmic strands and inclusion bodies in the cortical regions of the cell.
番茄黄化曲叶双生病毒以色列分离株(TYLCV-Is)是一种主要的番茄病原体,在新旧世界均造成大量作物损失。然而,令人惊讶的是,关于TYLCV-Is与番茄细胞相互作用的分子机制却知之甚少。在此,我们鉴定出一种TYLCV-Is蛋白V2,它作为RNA沉默抑制子,与目前已知的病毒抑制子无关。具体而言,是V2而非TYLCV-Is的其他蛋白抑制了报告转基因绿色荧光蛋白(GFP)的RNA沉默。这种抑制作用提高了GFP转录本和GFP蛋白的细胞水平,但对GFP特异性小干扰RNA(siRNA)的积累没有明显影响,这表明TYLCV-Is V2靶向RNA沉默途径中Dicer介导的双链RNA切割之后的一个步骤。对TYLCV-Is V2在植物原生质体和组织中的亚细胞定位进行可视化显示,该蛋白与细胞皮层区域的细胞质丝和内含体相关。