Localization of ASH1 mRNA particles in living yeast
Localization of ASH1 mRNA particles in living yeast
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DOI：
10.1016/s1097-2765(00)80143-4
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发表时间：
1998-10-01
期刊：
影响因子：
16
通讯作者：
Long, RM
中科院分区：
文献类型：
--
作者：
Bertrand, E;Chartrand, P;Long, RM
ASH 1 mRNA定位于酿酒酵母的芽尖，以建立HO表达的不对称性，这对交配型转换很重要。为了观察mRNA在活酵母细胞中的真实的时间定位，将绿色荧光蛋白（GFP）与RNA结合蛋白MS 2融合，以跟随含有MS 2结合位点的报告mRNA。GFP颗粒在芽中的形成和定位需要ASH 1 3 'UTR（非翻译区）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。She 3 myc和She 1 myc都与该粒子共域。视频显微镜显示She 1 p/Myo 4p以200-440 nm/sec的速度将颗粒移动到芽尖。因此，ASH 1 3 'UTR依赖性颗粒可作为RNA转运和定位的标志物。
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.