Bertrand E, Chartrand P, Schaefer M, Shenoy S M, Singer R H, Long R M
Department of Anatomy and Structural Biology and Cell Biology, Albert Einstein College of Medicine, Bronx, New York 10461, USA.
Mol Cell. 1998 Oct;2(4):437-45. doi: 10.1016/s1097-2765(00)80143-4.
ASH1 mRNA localizes to the bud tip in Saccharomyces cerevisiae to establish asymmetry of HO expression, important for mating type switching. To visualize real time localization of the mRNA in living yeast cells, green fluorescent protein (GFP) was fused to the RNA-binding protein MS2 to follow a reporter mRNA containing MS2-binding sites. Formation and localization of a GFP particle in the bud required ASH1 3'UTR (untranslated region) sequences. The SHE mutants disrupt RNA and particle localization and SHE 2 and 3 mutants inhibit particle formation as well. Both She3myc and She1myc colocalized with the particle. Video microscopy demonstrated that She1p/Myo4p moved particles to the bud tip at 200-440 nm/sec. Therefore, the ASH1 3'UTR-dependent particle serves as a marker for RNA transport and localization.
在酿酒酵母中，ASH1信使核糖核酸（mRNA）定位于芽尖，以建立HO表达的不对称性，这对于交配型转换很重要。为了在活酵母细胞中可视化mRNA的实时定位，绿色荧光蛋白（GFP）与RNA结合蛋白MS2融合，以追踪含有MS2结合位点的报告mRNA。芽中GFP颗粒的形成和定位需要ASH1 3'非翻译区（UTR）序列。SHE突变体破坏RNA和颗粒定位，SHE 2和3突变体也抑制颗粒形成。She3myc和She1myc都与颗粒共定位。视频显微镜显示，She1p/Myo4p以200 - 440纳米/秒的速度将颗粒移动到芽尖。因此，依赖ASH1 3'UTR的颗粒可作为RNA运输和定位的标志物。
