Leh V, Jacquot E, Geldreich A, Hermann T, Leclerc D, Cerutti M, Yot P, Keller M, Blanc S
Institut de Biologie Moléculaire des Plantes, UPR CNRS 406, 12 rue du Général Zimmer, 67084 Strasbourg Cedex.
EMBO J. 1999 Dec 15;18(24):7077-85. doi: 10.1093/emboj/18.24.7077.
The open reading frame (ORF) III product (PIII) of cauliflower mosaic virus is necessary for the infection cycle but its role is poorly understood. We have used in vitro protein binding ('far Western') assays to demonstrate that PIII interacts with the cauliflower mosaic virus (CaMV) ORF II product (PII), a known aphid transmission factor. Aphid transmission of purified virions of the PII-defective strain CM4-184 was dependent upon added PII, but complementation was efficient only in the presence of PIII, demonstrating the requirement of PIII for transmission. Deletion mutagenesis mapped the interaction domains of PIII and PII to the 30 N-terminal and 61 C-terminal residues of PIII and PII, respectively. A model for interaction between PIII and PII is proposed on the basis of secondary structure predictions. Finally, a direct correlation between the ability of PIII and PII to interact and aphid transmissibility of the virus was demonstrated by using mutagenized PIII proteins. Taken together, these data argue strongly that PIII is a second 'helper' factor required for CaMV transmission by aphids.
花椰菜花叶病毒的开放阅读框(ORF)III产物(PIII)是感染周期所必需的,但其作用尚不清楚。我们利用体外蛋白质结合(“远Western”)试验证明,PIII与花椰菜花叶病毒(CaMV)的ORF II产物(PII)相互作用,PII是一种已知的蚜虫传播因子。PII缺陷型菌株CM4-184的纯化病毒粒子的蚜虫传播依赖于添加的PII,但只有在PIII存在的情况下互补才有效,这表明PIII是传播所必需的。缺失诱变将PIII和PII的相互作用结构域分别定位到PIII的30个N端残基和PII的61个C端残基。基于二级结构预测提出了PIII和PII之间相互作用的模型。最后,通过使用诱变的PIII蛋白,证明了PIII和PII相互作用的能力与病毒的蚜虫传播性之间存在直接相关性。综上所述,这些数据有力地表明,PIII是蚜虫传播CaMV所需的第二个“辅助”因子。