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首页> 外文期刊>Protoplasma: An International Journal of Cell Biology >Intranuclear accumulation of plant tubulin in response to low temperature
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Intranuclear accumulation of plant tubulin in response to low temperature

机译：低温下植物微管蛋白的核内积累

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摘要

Concurrently with cold-induced disintegration of microtubular structures in the cytoplasm, gradual tubulin accumulation was observed in a progressively growing proportion of interphase nuclei in tobacco BY-2 cells. This intranuclear tubulin disappeared upon rewarming. Simultaneously, new micrombules rapidly emerged from the nuclear periphery and reconstituied new cortical arrays, as was shown by immunofluorescence. A rapid exclusion of tubulin from the nucleus during rewarming was also observed in vivo in cells expressing GFP-tubulin. Nuclei were purified from cells that expressed GFP fused to an endo-plastnic-reticulum retention signal (BY-2-mGFP5-ER), and green-fluorescent protein was used as a diagnostic marker to confirm that the nuclear fraction was not contaminated by nuclear-envelope proteins, These purified, GFP-free nuclei contained tubulin when isolated from cold-treated cells, whereas control nuclei were void of tubulin. Furthermore, highly conserved putative nuclear-export sequences were identified in tubulin sequences. These results led us to interpret the accumulation of tubulin in interphasic nuclei, as well as its rapid nuclear export, in the context of ancient intranuclear tubulin function during the cell cycle progression.
机译：在细胞质中冷诱导的微管结构崩解的同时，在烟草BY-2细胞中，相间核的比例逐渐增加，观察到微管蛋白逐渐积累。核内微管蛋白在重新温热后消失。同时，如免疫荧光所显示，新的微粒子从核外围迅速出现并重构了新的皮层阵列。在体内，在表达GFP-微管蛋白的细胞中也观察到微管蛋白从核中快速排除在细胞核之外。从表达融合有内质网保留信号（BY-2-mGFP5-ER）的GFP的细胞中纯化细胞核，并使用绿色荧光蛋白作为诊断标记，以确认核级分未被核污染-包膜蛋白，当从冷处理的细胞中分离时，这些纯化的，不含GFP的细胞核中含有微管蛋白，而对照细胞核中没有微管蛋白。此外，在微管蛋白序列中鉴定出高度保守的推定核输出序列。这些结果使我们在细胞周期进程中古老的核内微管蛋白功能的背景下解释了微管蛋白在相间核中的积累及其快速的核输出。

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