Yelina Natalia E, Savenkov Eugene I, Solovyev Andrey G, Morozov Sergey Y, Valkonen Jari P T
Department of Virology and A. N. Belozersky Institute of Physico-Chemical Biology, Moscow State University, Moscow 119899, Russia.
J Virol. 2002 Dec;76(24):12981-91. doi: 10.1128/jvi.76.24.12981-12991.2002.
RNA silencing is a natural defense mechanism against genetic stress factors, including viruses. A mutant hordeivirus (Barley stripe mosaic virus [BSMV]) lacking the gammab gene was confined to inoculated leaves in Nicotiana benthamiana, but systemic infection was observed in transgenic N. benthamiana expressing the potyviral silencing suppressor protein HCpro, suggesting that the gammab protein may be a long-distance movement factor and have antisilencing activity. This was shown for gammab proteins of both BSMV and Poa semilatent virus (PSLV), a related hordeivirus. Besides the functions in RNA silencing suppression, gammab and HCpro had analogous effects on symptoms induced by the hordeiviruses. Severe BSMV-induced symptoms were correlated with high HCpro concentrations in the HCpro-transgenic plants, and substitution of the gammab cistron of BSMV with that of PSLV led to greatly increased symptom severity and an altered pattern of viral gene expression. The efficient systemic infection with the chimera was followed by the development of dark green islands (localized recovery from infection) in leaves and exemption of new developing leaves from infection. Recovery and the accumulation of short RNAs diagnostic of RNA silencing in the recovered tissues in wild-type N. benthamiana were suppressed in HCpro-transgenic plants. These results provide evidence that potyviral HCpro and hordeivirus gammab proteins contribute to systemic viral infection, symptom severity, and RNA silencing suppression. HCpro's ability to suppress the recovery of plants from viral infection emphasizes recovery as a manifestation of RNA silencing.
RNA沉默是一种针对包括病毒在内的遗传应激因素的天然防御机制。一种缺失γb基因的突变大麦病毒(大麦条纹花叶病毒[BSMV])局限于本氏烟草的接种叶片中,但在表达马铃薯Y病毒属沉默抑制蛋白HCpro的转基因本氏烟草中观察到了系统感染,这表明γb蛋白可能是一种长距离移动因子并具有抗沉默活性。BSMV和Poa半潜病毒(PSLV,一种相关的大麦病毒)的γb蛋白均显示出这种特性。除了在RNA沉默抑制中的功能外,γb和HCpro对大麦病毒诱导的症状具有类似的影响。在HCpro转基因植物中,严重的BSMV诱导症状与高浓度的HCpro相关,用PSLV的γb顺反子替换BSMV的γb顺反子导致症状严重程度大幅增加以及病毒基因表达模式改变。用嵌合体进行的有效系统感染之后,叶片中出现深绿色岛屿(感染的局部恢复),并且新长出的叶片免于感染。在野生型本氏烟草中,恢复以及在恢复组织中出现诊断RNA沉默的短RNA积累在HCpro转基因植物中受到抑制。这些结果提供了证据,表明马铃薯Y病毒属HCpro和大麦病毒γb蛋白有助于病毒的系统感染、症状严重程度以及RNA沉默抑制。HCpro抑制植物从病毒感染中恢复的能力强调了恢复是RNA沉默的一种表现。