Simultaneous visualization of peroxisomes and cytoskeletal elements reveals actin and not microtubule-based peroxisome motility in plants
Simultaneous visualization of peroxisomes and cytoskeletal elements reveals actin and not microtubule-based peroxisome motility in plants
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DOI：
10.1104/pp.011018
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发表时间：
2002-03-01
期刊：
影响因子：
7.4
通讯作者：
Hülskamp, M
中科院分区：
文献类型：
--
作者：
Mathur, J;Mathur, N;Hülskamp, M
过氧化物酶体在活的植物细胞中使用标记有由SKL基序组成的过氧化物酶体靶向信号的黄色荧光蛋白可视化。过氧化物酶体和微丝/微管的同时可视化完成洋葱（洋葱）表皮细胞瞬时表达的黄色荧光蛋白peroxi结构，一个绿色荧光蛋白mTalin结构，标记的粘附肌动蛋白丝，和一个绿色荧光蛋白微管结合结构域的结构，标记微管。过氧化物酶体和细胞骨架元件的共可视化显示，与动物细胞的报道相反，植物中的过氧化物酶体似乎与肌动蛋白丝而不是微管相关。药理学研究表明，过氧化物酶体运动是基于肌动蛋白的。对于这个分析，我们使用洋葱表皮细胞和各种细胞类型的拟南芥，包括毛状体，根毛，根皮层细胞表现出不同的生长模式。在瞬时洋葱表皮测定和转基因拟南芥植物中，肌动蛋白细胞骨架的干扰导致跳跃运动的逐渐丧失，随后在30分钟内的药物应用的聚集和过氧化物酶体运动的完全停止。微管解聚或稳定没有影响。
Peroxisomes were visualized in living plant cells using a yellow fluorescent protein tagged with a peroxisomal targeting signal consisting of the SKL motif. Simultaneous visualization of peroxisomes and microfilaments/microtubules was accomplished in onion (Allium cepa) epidermal cells transiently expressing the yellow fluorescent protein-peroxi construct, a green fluorescent protein-mTalin construct that labels filamentous-actin filaments, and a green fluorescent proteinmicrotubule-binding domain construct that labels microtubules. The covisualization of peroxisomes and cytoskeletal elements revealed that, contrary to the reports from animal cells, peroxisomes in plants appear to associate with actin filaments and not microtubules, That peroxisome movement is actin based was shown by pharmacological studies. For this analysis we used onion epidermal cells and various cell types of Arabidopsis including trichomes, root hairs, and root cortex cells exhibiting different modes of growth. In transient onion epidermis assay and in transgenic Arabidopsis plants, an interference with the actin cytoskeleton resulted in progressive loss of saltatory movement followed by the aggregation and a complete cessation of peroxisome motility within 30 min of drug application. Microtubule depolymerization or stabilization had no effect.