Bucher Etienne, Sijen Titia, De Haan Peter, Goldbach Rob, Prins Marcel
Laboratory of Virology, Wageningen University, The Netherlands.
J Virol. 2003 Jan;77(2):1329-36. doi: 10.1128/jvi.77.2.1329-1336.2003.
Posttranscriptional silencing of a green fluorescent protein (GFP) transgene in Nicotiana benthamiana plants was suppressed when these plants were infected with Tomato spotted wilt virus (TSWV), a plant-infecting member of the BUNYAVIRIDAE: Infection with TSWV resulted in complete reactivation of GFP expression, similar to the case for Potato virus Y, but distinct from that for Cucumber mosaic virus, two viruses known to carry genes encoding silencing suppressor proteins. Agrobacterium-based leaf injections with individual TSWV genes identified the NS(S) gene to be responsible for the RNA silencing-suppressing activity displayed by this virus. The absence of short interfering RNAs in NS(S)-expressing leaf sectors suggests that the tospoviral NS(S) protein interferes with the intrinsic RNA silencing present in plants. Suppression of RNA silencing was also observed when the NS3 protein of the Rice hoja blanca tenuivirus, a nonenveloped negative-strand virus, was expressed. These results indicate that plant-infecting negative-strand RNA viruses carry a gene for a suppressor of RNA silencing.
当本氏烟草植株感染番茄斑萎病毒(TSWV,布尼亚病毒科的一种植物感染病毒)时,绿色荧光蛋白(GFP)转基因在这些植株中的转录后沉默被抑制:感染TSWV导致GFP表达完全重新激活,这与马铃薯Y病毒的情况类似,但与黄瓜花叶病毒不同,后两种病毒已知携带编码沉默抑制蛋白的基因。用单个TSWV基因进行基于农杆菌的叶片注射,确定NS(S)基因负责该病毒所表现出的RNA沉默抑制活性。在表达NS(S)的叶片区域中不存在短干扰RNA,这表明番茄斑萎病毒NS(S)蛋白干扰了植物中固有的RNA沉默。当表达水稻白叶病毒(一种无包膜负链病毒)的NS3蛋白时,也观察到了RNA沉默的抑制。这些结果表明,植物感染性负链RNA病毒携带一个RNA沉默抑制子基因。