Gao Le, Tuo Decai, Shen Wentao, Yan Pu, Li Xiaoying, Zhou Peng
Key Laboratory of Biology and Genetic Resources of Tropical Crops, Ministry of Agriculture, Institute of Tropical Bioscience and Biotechnology & Analysis and Testing Center, Chinese Academy of Tropical Agricultural Sciences, Haikou, 571101, People's Republic of China.
Virus Genes. 2015 Feb;50(1):97-103. doi: 10.1007/s11262-014-1145-x. Epub 2014 Nov 22.
The interaction of papaya eukaryotic translation initiation factor 3 subunit G (CpeIF3G) with Papaya ringspot virus (PRSV) NIa-Pro was validated using a bimolecular fluorescence complementation assay in papaya protoplasts based on the previous yeast two-hybrid assay results. The C-terminal (residues 133-239) fragment of PRSV NIa-Pro and the central domain (residues 59-167) of CpeIF3G were required for effective interaction between NIa-Pro and CpeIF3G as shown by a Sos recruitment yeast two-hybrid system with several deletion mutants of NIa-Pro and CpeIF3G. The central domain of CpeIF3G, which contains a C2HC-type zinc finger motif, is required to bind to other eIFs of the translational machinery. In addition, quantitative real-time reverse transcription PCR assay confirmed that PRSV infection leads to a 2- to 4.5-fold up-regulation of CpeIF3G mRNA in papaya. Plant eIF3G is involved in various stress response by enhancing the translation of resistance-related proteins. It is proposed that the NIa-Pro-CpeIF3G interaction may impair translation preinitiation complex assembly of defense proteins and interfere with host defense.
基于先前的酵母双杂交实验结果，在木瓜原生质体中利用双分子荧光互补实验验证了木瓜真核翻译起始因子3亚基G（CpeIF3G）与木瓜环斑病毒（PRSV）NIa-Pro之间的相互作用。Sos募集酵母双杂交系统对NIa-Pro和CpeIF3G的多个缺失突变体进行检测，结果表明，PRSV NIa-Pro的C末端（第133 - 239位氨基酸残基）片段和CpeIF3G的中央结构域（第59 - 167位氨基酸残基）是NIa-Pro与CpeIF3G有效相互作用所必需的。CpeIF3G的中央结构域含有一个C2HC型锌指基序，该结构域对于与翻译机制中的其他真核翻译起始因子（eIFs）结合是必需的。此外，实时定量逆转录PCR实验证实，PRSV感染导致木瓜中CpeIF3G mRNA上调2至4.5倍。植物eIF3G通过增强抗性相关蛋白的翻译参与多种应激反应。有人提出，NIa-Pro与CpeIF3G的相互作用可能会损害防御蛋白翻译起始前复合物的组装，并干扰宿主防御。
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