Jiang Yi, Serviene Elena, Gal Jozsef, Panavas Tadas, Nagy Peter D
Department of Plant Pathology, University of Kentucky, Plant Science Building, Lexington, KY 40546, USA.
J Virol. 2006 Aug;80(15):7394-404. doi: 10.1128/JVI.02686-05.
To identify essential host genes affecting replication of Tomato bushy stunt virus (TBSV), a small model plant virus, we screened 800 yeast genes present in the yeast Tet promoters Hughes Collection. In total, we have identified 30 new host genes whose down-regulation either increased or decreased the accumulation of a TBSV replicon RNA. The identified essential yeast genes are involved in RNA transcription/metabolism, protein metabolism/transport, or other cellular processes. Detailed analysis of the effects of some of the identified yeast genes revealed that they might affect RNA replication by altering (i) the amounts/functions of p33 and p92(pol) viral replication proteins, (ii) the standard 10 to 20:1 ratio between p33 and p92(pol) in the viral replicase, (iii) the activity of the tombusvirus replicase, and (iv) the ratio of plus- versus minus-stranded RNA replication products. Altogether, this and previous genetic screening of yeast (Panavas et al., Proc. Natl. Acad. Sci. USA 102:7326-7331, 2005) led to the identification of 126 host genes (out of approximately 5,600 genes that represent approximately 95% of all the known and predicted yeast genes) that affected the accumulation of tombusvirus RNA.
为了鉴定影响小型模式植物病毒番茄丛矮病毒（TBSV）复制的宿主必需基因，我们对酵母Tet启动子休斯文库中存在的800个酵母基因进行了筛选。我们总共鉴定出30个新的宿主基因，其下调会增加或减少TBSV复制子RNA的积累。所鉴定出的酵母必需基因参与RNA转录/代谢、蛋白质代谢/运输或其他细胞过程。对一些已鉴定出的酵母基因作用的详细分析表明，它们可能通过改变以下方面来影响RNA复制：（i）p33和p92（pol）病毒复制蛋白的量/功能；（ii）病毒复制酶中p33和p92（pol）之间标准的10比20至1的比例；（iii）番茄病毒复制酶的活性；（iv）正链与负链RNA复制产物的比例。总之，本次研究以及之前对酵母的遗传筛选（帕纳瓦斯等人，《美国国家科学院院刊》102:7326 - 7331，2005年）导致鉴定出126个影响番茄病毒RNA积累的宿主基因（在大约5600个基因中，这些基因约占所有已知和预测酵母基因的95%）。
