Saito T, Hosokawa D, Meshi T, Okada Y
Department of Biophysics and Biochemistry, Faculty of Science, University of Tokyo, Hongo, Tokyo 113, Japan.
Virology. 1987 Oct;160(2):477-81. doi: 10.1016/0042-6822(87)90020-1.
We have revealed the cellular localization of the putative replicase components of tobacco mosaic virus (TMV), 130K and 180K proteins, in TMV-infected tobacco leaves by the immunogold technique with antisera which specifically react with these two proteins. When sections of TMV-infected tobacco leaves were treated with anti-130K protein antiserum and then with protein A-gold complex, most of the gold label was strongly localized on granular inclusion bodies which were found specifically in the cytoplasm of TMV-infected cells. Very small amounts of label present in other regions, including the nuclei, chloroplasts, and mitochondria, seemed to be nonspecific. Gold-labeled 180K protein was also dispersed over the granular inclusion bodies. The granular inclusion bodies appeared to be oval-shaped structures with various diameters ranging from 0.2 to 2.8 microm. TMV particles were usually observed near the granular inclusion bodies as aggregates but not inside them. Considering the involvement of the 130K and 180K proteins in replication, the granular inclusion bodies may be the site for replication of TMV RNA.
我们通过免疫金技术,利用能与烟草花叶病毒(TMV)的130K和180K蛋白特异性反应的抗血清,揭示了TMV假定复制酶组分130K和180K蛋白在TMV感染的烟草叶片中的细胞定位。用抗130K蛋白抗血清处理TMV感染的烟草叶片切片,然后用蛋白A-金复合物处理,大部分金标记强烈定位于颗粒状内含体上,这些内含体专门存在于TMV感染细胞的细胞质中。在包括细胞核、叶绿体和线粒体在内的其他区域存在的极少量标记似乎是非特异性的。金标记的180K蛋白也分散在颗粒状内含体上。颗粒状内含体似乎是椭圆形结构,直径从0.2到2.8微米不等。TMV颗粒通常在颗粒状内含体附近以聚集体形式观察到,但不在其内部。考虑到130K和180K蛋白参与复制,颗粒状内含体可能是TMV RNA复制的场所。