Kim Min Jung, Huh Sung Un, Ham Byung-Kook, Paek Kyung-Hee
School of Life Sciences and Biotechnology, Korea University, 1, 5-ga, Anam-dong, Sungbuk-gu, Seoul 136-701, Republic of South Korea.
J Virol. 2008 May;82(10):4823-33. doi: 10.1128/JVI.02518-07. Epub 2008 Mar 5.
In mammalian and yeast systems, methyltransferases have been implicated in the regulation of diverse processes, such as protein-protein interactions, protein localization, signal transduction, RNA processing, and transcription. The Cucumber mosaic virus (CMV) 1a protein is essential not only for virus replication but also for movement. Using a yeast two-hybrid system with tobacco plants, we have identified a novel gene encoding a methyltransferase that interacts with the CMV 1a protein and have designated this gene Tcoi1 (tobacco CMV 1a-interacting protein 1). Tcoi1 specifically interacted with the methyltransferase domain of CMV 1a, and the expression of Tcoi1 was increased by CMV inoculation. Biochemical studies revealed that the interaction of Tcoi1 with CMV 1a protein was direct and that Tcoi1 methylated CMV 1a protein both in vitro and in vivo. The CMV 1a binding activity of Tcoi1 is in the C-terminal domain, which shows the methyltransferase activity. The overexpression of Tcoi1 enhanced the CMV infection, while the reduced expression of Tcoi1 decreased virus infectivity. These results suggest that Tcoi1 controls the propagation of CMV through an interaction with the CMV 1a protein.
在哺乳动物和酵母系统中,甲基转移酶参与多种过程的调控,如蛋白质-蛋白质相互作用、蛋白质定位、信号转导、RNA加工和转录。黄瓜花叶病毒(CMV)1a蛋白不仅对病毒复制至关重要,对病毒移动也很关键。利用烟草植物的酵母双杂交系统,我们鉴定出一个编码与CMV 1a蛋白相互作用的甲基转移酶的新基因,并将其命名为Tcoi1(烟草CMV 1a相互作用蛋白1)。Tcoi1特异性地与CMV 1a的甲基转移酶结构域相互作用,CMV接种可使Tcoi1的表达增加。生化研究表明,Tcoi1与CMV 1a蛋白的相互作用是直接的,且Tcoi1在体外和体内均可使CMV 1a蛋白甲基化。Tcoi1的CMV 1a结合活性位于C末端结构域,该结构域具有甲基转移酶活性。Tcoi1的过表达增强了CMV感染,而Tcoi1表达降低则降低了病毒感染性。这些结果表明,Tcoi1通过与CMV 1a蛋白相互作用来控制CMV的传播。