Golgi membrane dynamics after induction of a dominant-negative mutant Sar1 GTPase in tobacco.
Golgi membrane dynamics after induction of a dominant-negative mutant Sar1 GTPase in tobacco.
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DOI：
10.1093/jxb/erp315
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发表时间：
2010
影响因子：
6.9
通讯作者：
A. Osterrieder;E. Hummel;Claudine M Carvalho;C. Hawes
中科院分区：
文献类型：
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作者：
A. Osterrieder;E. Hummel;Claudine M Carvalho;C. Hawes
在烟草植物中建立了一个可诱导的系统，允许控制Sar 1-GTP的表达，从而研究抑制内质网（ER）向高尔基体转运后的蛋白质动力学。在诱导后18- 24小时内观察到高尔基体完全解体和高尔基体标记重新分布到ER中。在超微结构水平上，Sar 1-GTP的表达导致高尔基体堆叠大小减少，随后是高尔基体碎片化和囊泡残余物的积累。Sar 1-GTP的诱导导致绿色荧光蛋白（GFP）标记的拟南芥golgin AtCASP和GC 1（golgin候选物1，拟南芥golgin 84同种型）分别重新分布到ER或细胞质中。此外，这两种融合蛋白在点状结构中观察到，其与黄色荧光蛋白（YFP）标记的Sar 1-GTP版本共定位。Sar 1-GTP诱导系统进行了比较组成Sar 1-GTP表达和布雷菲德菌素A治疗，并讨论了其潜在的ER出口位点和早期顺式高尔基体结构的组成的研究。
An inducible system has been established in Nicotiana tabacum plants allowing controlled expression of Sar1-GTP and thus the investigation of protein dynamics after inhibition of endoplasmic reticulum (ER) to Golgi transport. Complete Golgi disassembly and redistribution of Golgi markers into the ER was observed within 18-24h after induction. At the ultrastructural level Sar1-GTP expression led to a decrease in Golgi stack size followed by Golgi fragmentation and accumulation of vesicle remnants. Induction of Sar1-GTP resulted in redistribution of the green fluorescent protein (GFP)-tagged Arabidopsis golgins AtCASP and GC1 (golgin candidate 1, an Arabidopsis golgin 84 isoform) into the ER or cytoplasm, respectively. Additionally, both fusion proteins were observed in punctate structures, which co-located with a yellow fluorescent protein (YFP)-tagged version of Sar1-GTP. The Sar1-GTP-inducible system is compared with constitutive Sar1-GTP expression and brefeldin A treatment, and its potential for the study of the composition of ER exit sites and early cis-Golgi structures is discussed.