A role for clathrin in reassembly of the golgi apparatus
A role for clathrin in reassembly of the golgi apparatus
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DOI：
10.1091/mbc.e06-06-0532
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发表时间：
2007-01-01
影响因子：
3.3
通讯作者：
Shields, Dennis
中科院分区：
文献类型：
--
作者：
Radulescu, Andreea E.;Siddhanta, Anirban;Shields, Dennis
高尔基体是一个高度动态的细胞器，其组织由蛋白质基质、细胞骨架成分和肌醇磷脂维持。在哺乳动物细胞中，细胞器的解体发生在有丝分裂开始时是可逆的，而在细胞凋亡时是不可逆的。几种药物包括诺可达唑、布雷菲尔丁素A （BFA）和伯醇（1-丁醇）可诱导高尔基体可逆断裂。为了研究高尔基重组的机制，我们用1-丁醇、BFA或诺可达唑处理大鼠NRK和GH3细胞。在1-丁醇洗脱过程中，网格蛋白（一种普遍存在的参与跨高尔基网络和质膜囊泡运输的外壳蛋白）和大量的网格蛋白包被囊泡被招募到新生高尔基池区域。内源性网格蛋白重链的敲低表明去除BFA或1-丁醇后高尔基体无法有效重组。相反，在1-丁醇冲洗后，它保持致密，紧密的形态。我们的研究结果表明，网格蛋白是重组碎片化高尔基体元素所必需的。此外，我们表明，经过丁醇处理后，高尔基体通过最初的致密中间结构进行改革，随后重塑为特征的间相网状形态，重新组装需要网格蛋白。
The Golgi apparatus is a highly dynamic organelle whose organization is maintained by a proteinaceous matrix, cytoskeletal components, and inositol phospholipids. in mammalian cells, disassembly of the organelle occurs reversibly at the onset of mitosis and irreversibly during apoptosis. Several pharmacological agents including nocodazole, brefeldin A (BFA), and primary alcohols (1-butanol) induce reversible fragmentation of the Golgi apparatus. To dissect the mechanism of Golgi reassembly, rat NRK and GH3 cells were treated with 1-butanol, BFA, or nocodazole. During washout of 1-butanol, clathrin, a ubiquitous coat protein implicated in vesicle traffic at the trans-Golgi network and plasma membrane, and abundant clathrin coated vesicles were recruited to the region of nascent Golgi cisternae. Knockdown of endogenous clathrin heavy chain showed that the Golgi apparatus failed to reform efficiently after BFA or 1-butanol removal. Instead, upon 1-butanol washout, it maintained a compact, tight morphology. Our results suggest that clathrin is required to reassemble fragmented Golgi elements. In addition, we show that after butanol treatment the Golgi apparatus reforms via an initial compact intermediate structure that is subsequently remodeled into the characteristic interphase lace-like morphology and that reassembly requires clathrin.