Sequential depletion and acquisition of proteins during Golgi stack disassembly and reformation.
Sequential depletion and acquisition of proteins during Golgi stack disassembly and reformation.
复制标题
高尔基体堆叠拆卸和重组过程中蛋白质的连续消耗和获取。
DOI：
10.1111/j.1600-0854.2010.01106.x
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发表时间：
2010
期刊：
影响因子：
--
通讯作者：
Schoberer J
中科院分区：
文献类型：
--
作者：
Schoberer J
在此，我们报道了在GTP锁定的Sar1p或Brefeldin A(BFA)的诱导表达和BFA洗脱时，多种植物高尔基膜标志物在烟草叶表皮细胞高尔基体解体过程中的逐步转运和重组。在拆解和重新组装过程中，通过共聚焦显微镜跟踪具有特定顺式-反式高尔基体亚位置的荧光高尔基体N-葡聚糖加工酶和基质蛋白(高尔基体)的分布。高尔基体分解过程中的第一个事件是高尔基体膜上的反式高尔基体酶和高尔基体蛋白的丢失，随后中间和顺式高尔基体酶重新分布到内质网(ER)，而高尔基体被重新定位到内质网或细胞质。分离纯化和免疫印迹证实了该蛋白的存在。高尔基体组分在反式-顺式序列中的顺序重新分布可能突出了植物中反式-高尔基体与内质网之间的一条新的逆行运输途径。BFA洗涤后，高尔基体标志物从内质网中以相反的顺序释放，顺式基质蛋白标记高尔基体结构先于肌层/中层酶。在跨酶定位之前，反式基质蛋白被招募回高尔基体膜。我们的结果表明，高尔基体的拆解和重组在植物中以高度有序的方式发生。
Herein, we report the stepwise transport of multiple plant Golgi membrane markers during disassembly of the Golgi apparatus in tobacco leaf epidermal cells in response to the induced expression of the GTP‐locked Sar1p or Brefeldin A (BFA), and reassembly on BFA washout. The distribution of fluorescent Golgi‐residentN‐glycan processing enzymes and matrix proteins (golgins) with specificcis–trans‐Golgi sub‐locations was followed by confocal microscopy during disassembly and reassembly. The first event during Golgi disassembly was the loss oftrans‐Golgi enzymes and golgins from Golgi membranes, followed by a sequential redistribution of medial andcis‐Golgi enzymes into the endoplasmic reticulum (ER), whilst golgins were relocated to the ER or cytoplasm. This event was confirmed by fractionation and immuno‐blotting. The sequential redistribution of Golgi components in atrans–cissequence may highlight a novel retrograde trafficking pathway between thetrans‐Golgi and the ER in plants. Release of Golgi markers from the ER upon BFA washout occurred in the opposite sequence, withcis‐matrix proteins labelling Golgi‐like structures beforecis/medial enzymes.Trans‐enzyme location was preceded bytrans‐matrix proteins being recruited back to Golgi membranes. Our results show that Golgi disassembly and reassembly occur in a highly ordered fashion in plants.