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首页> 外文期刊>Protoplasma: An International Journal of Cell Biology >Disturbance of endomembrane trafficking by brefeldin A and calyculin A reorganizes the actin cytoskeleton of Lilium longiflorum pollen tubes
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Disturbance of endomembrane trafficking by brefeldin A and calyculin A reorganizes the actin cytoskeleton of Lilium longiflorum pollen tubes

机译：布雷菲德菌素A和calyculin A对内膜运输的干扰重新组织了百合花粉花粉管的肌动蛋白细胞骨架

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摘要

We investigated the effect of brefeldin A on membrane trafficking and the actin cytoskeleton of pollen tubes of Lilium longiflorum with fluorescent dyes, inhibitor experiments, and confocal laser scanning microscopy. The formation of a subapical brefeldin A-induced membrane aggregation (BIA) was associated with the formation of an actin basket from which filaments extended towards the tip. The orientation of these actin filaments correlated with the trajectories of membrane material stained by FM dyes, suggesting that the BIA-associated actin filaments are used as tracks for retrograde transport. Analysis of time series indicated that these tracks ( actin filaments) were either stationary or glided along the plasma membrane towards the BIA together with the attached membranes or organelles. Disturbance of the actin cytoskeleton by cytochalasin D or latrunculin B caused immediate arrest of membrane trafficking, dissipation of the BIA and the BIA-associated actin basket, and reorganization into randomly oriented actin rods. Our observations suggest that brefeldin A causes ectopic activation of actin-nucleating proteins at the BIA, resulting in retrograde movement of membranes not only along but also together with actin filaments. We show further that subapical membrane aggregations and actin baskets supporting retrograde membrane flow can also be induced by calyculin A, indicating that dephosphorylation by type 2 protein phosphatases is required for proper formation of membrane coats and polar membrane trafficking.
机译：我们用荧光染料，抑制剂实验和共聚焦激光扫描显微镜研究了布雷菲德菌素A对长花百合花粉管膜运输和肌动蛋白细胞骨架的影响。根尖下布雷菲德菌素A诱导的膜聚集（BIA）的形成与肌动蛋白篮的形成有关，细丝从肌动篮向尖端延伸。这些肌动蛋白丝的取向与被FM染料染色的膜材料的轨迹相关，这表明与BIA相关的肌动蛋白丝被用作逆行转运的轨道。时间序列分析表明，这些径迹（肌动蛋白丝）是固定的，或者沿着质膜与附着的膜或细胞器一起向BIA滑动。细胞松弛素D或latrunculin B对肌动蛋白细胞骨架的干扰会立即阻止膜运输，消散BIA和与BIA相关的肌动蛋白篮，并重组为随机定向的肌动蛋白棒。我们的观察结果表明，布雷菲德菌素A在BIA处引起肌动蛋白成核蛋白的异位活化，导致膜不仅沿肌动蛋白丝而且与肌动蛋白丝一起逆行运动。我们进一步表明，顶叶下膜聚集和支持逆行膜流的肌动蛋白篮也可以被calyculin A诱导，表明通过2型蛋白磷酸酶的去磷酸化是正确形成膜衣和极性膜运输所必需的。

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