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Living on the edge: the role of Atgolgin‐84A at the plant ER‐Golgi interface
Journal of Microscopy ( IF 1.5 ) Pub Date : 2020-07-31 , DOI: 10.1111/jmi.12946
V Vieira 1, 2 , C Pain 1 , S Wojcik 1 , T Spatola Rossi 1 , J Denecke 3 , A Osterrieder 1, 4 , C Hawes 1 , V Kriechbaumer 1
Affiliation  

The plant Golgi apparatus is responsible for the processing of proteins received from the endoplasmic reticulum (ER) and their distribution to multiple destinations within the cell. Golgi matrix components, such as golgins, have been identified and suggested to function as putative tethering factors to mediate the physical connections between Golgi bodies and the ER network. Golgins are proteins anchored to the Golgi membrane by the C‐terminus either through transmembrane domains or interaction with small regulatory GTPases. The golgin N‐terminus contains long coiled‐coil domains, which consist of a number of α‐helices wrapped around each other to form a structure similar to a rope being made from several strands, reaching into the cytoplasm. In animal cells, golgins are also implicated in specific recognition of cargo at the Golgi.Here, we investigate the plant golgin Atgolgin‐84A for its subcellular localization and potential role as a tethering factor at the ER–Golgi interface. For this, fluorescent fusions of Atgolgin‐84A and an Atgolgin‐84A truncation lacking the coiled‐coil domains (Atgolgin‐84AΔ1‐557) were transiently expressed in tobacco leaf epidermal cells and imaged using high‐resolution confocal microscopy. We show that Atgolgin‐84A localizes to a pre‐cis‐Golgi compartment that is also labelled by one of the COPII proteins as well as by the tether protein AtCASP. Upon overexpression of Atgolgin‐84A or its deletion mutant, transport between the ER and Golgi bodies is impaired and cargo proteins are redirected to the vacuole.

中文翻译:

生活在边缘:Atgolgin-84A 在植物 ER-Golgi 界面中的作用

植物高尔基体负责处理从内质网 (ER) 接收的蛋白质,并将它们分配到细胞内的多个目的地。高尔基体基质成分,如高尔金蛋白,已被确定并建议作为推定的束缚因子来调节高尔基体和 ER 网络之间的物理连接。高尔基蛋白是通过跨膜结构域或与小的调节性 GTP 酶相互作用通过 C 端锚定在高尔基体膜上的蛋白质。高尔金 N 端包含长卷曲螺旋结构域,由许多相互缠绕的 α 螺旋组成,形成类似于由几股组成的绳索的结构,伸入细胞质。在动物细胞中,高尔基体也与高尔基体的货物特异性识别有关。我们研究了植物高尔金 Atgolgin-84A 的亚细胞定位和作为内质网-高尔基体界面束缚因子的潜在作用。为此,Atgolgin-84A 和缺乏卷曲螺旋结构域的 Atgolgin-84A 截断(Atgolgin-84AΔ1-557)的荧光融合在烟叶表皮细胞中瞬时表达,并使用高分辨率共聚焦显微镜成像。我们表明 Atgolgin-84A 定位于一个 pre-cis-Golgi 区室,该区室也被 COPII 蛋白之一以及系链蛋白 AtCASP 标记。Atgolgin-84A 或其缺失突变体过表达后,内质网和高尔基体之间的运输受损,货物蛋白被重定向到液泡。Atgolgin-84A 和缺乏卷曲螺旋结构域的 Atgolgin-84A 截断(Atgolgin-84AΔ1-557)的荧光融合在烟叶表皮细胞中瞬时表达,并使用高分辨率共聚焦显微镜成像。我们表明 Atgolgin-84A 定位于一个 pre-cis-Golgi 区室,该区室也被 COPII 蛋白之一以及系链蛋白 AtCASP 标记。Atgolgin-84A 或其缺失突变体过表达后,内质网和高尔基体之间的运输受损,货物蛋白被重定向到液泡。Atgolgin-84A 和缺乏卷曲螺旋结构域的 Atgolgin-84A 截断(Atgolgin-84AΔ1-557)的荧光融合在烟叶表皮细胞中瞬时表达,并使用高分辨率共聚焦显微镜成像。我们表明 Atgolgin-84A 定位于一个 pre-cis-Golgi 区室,该区室也被 COPII 蛋白之一以及系链蛋白 AtCASP 标记。Atgolgin-84A 或其缺失突变体过表达后,内质网和高尔基体之间的运输受损,货物蛋白被重定向到液泡。我们表明 Atgolgin-84A 定位于一个 pre-cis-Golgi 区室,该区室也被 COPII 蛋白之一以及系链蛋白 AtCASP 标记。Atgolgin-84A 或其缺失突变体过表达后,内质网和高尔基体之间的运输受损,货物蛋白被重定向到液泡。我们表明 Atgolgin-84A 定位于一个 pre-cis-Golgi 区室,该区室也被 COPII 蛋白之一以及系链蛋白 AtCASP 标记。Atgolgin-84A 或其缺失突变体过表达后,内质网和高尔基体之间的运输受损,货物蛋白被重定向到液泡。
更新日期:2020-07-31
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