Mascarenhas J, Weber M H, Graumann P L
Biochemie, Fachbereich Chemie, Hans-Meerwein-Strasse, Philipps-Universität Marburg, 35032 Marburg, Germany.
EMBO Rep. 2001 Aug;2(8):685-9. doi: 10.1093/embo-reports/kve160. Epub 2001 Jul 19.
The large subunit of ribosomes in Bacillus subtilis was tagged by generation of a fusion of ribosomal protein L1 to blue fluorescent protein (BFP). The fusion was fully active and localized around the nucleoids, predominantly close to the cell poles, in growing cells. However, in stationary phase cells, and in growing cells treated with rifampicin, L1-BFP was distributed throughout the cells, in contrast to cells treated with chloramphenicol, in which ribosomes still localized around nucleoids. These data show that specific localization of ribosomes is not due to nucleoid exclusion, but is a dynamic process due to active synthesis of RNA. Dual labelling of ribosomes and cold shock proteins (CSPs) tagged with green fluorescent protein (GFP) revealed colocalization of both protein classes. CSPs are implicated in coupling of transcription with translation and may bridge the spatial separation of ribosomes and nucleoid-associated RNA polymerase.
通过将核糖体蛋白L1与蓝色荧光蛋白(BFP)融合,对枯草芽孢杆菌核糖体的大亚基进行了标记。该融合蛋白具有完全活性,在生长中的细胞中定位于类核周围,主要靠近细胞两极。然而,在稳定期细胞以及用利福平处理的生长细胞中,L1-BFP分布于整个细胞,这与用氯霉素处理的细胞形成对比,在用氯霉素处理的细胞中核糖体仍定位于类核周围。这些数据表明,核糖体的特异性定位并非由于类核排斥,而是由于RNA的活跃合成导致的一个动态过程。对核糖体和标记有绿色荧光蛋白(GFP)的冷休克蛋白(CSP)进行双重标记,结果显示这两类蛋白共定位。CSP与转录和翻译的偶联有关,可能在核糖体与类核相关的RNA聚合酶的空间分离之间起到桥梁作用。