Knockout of RSN1, TVP18 or CSC1‐2 causes perturbation of Golgi cisternae in Pichia pastoris,Traffic

当前位置： X-MOL 学术 › Traffic › 论文详情

Our official English website, www.x-mol.net, welcomes your feedback! (Note: you will need to create a separate account there.)

Knockout of RSN1, TVP18 or CSC1‐2 causes perturbation of Golgi cisternae in Pichia pastoris
Traffic ( IF 3.6 ) Pub Date : 2020-12-02 , DOI: 10.1111/tra.12773
Rochelle Aw _{1,

2} , Charlot De Wachter _{3,

4} , Bram Laukens _{3,

4} , Riet De Rycke _{5,

6} , Michiel De Bruyne _{5,

6} , David Bell _{7,

8} , Nico Callewaert _{3,

4} , Karen M. Polizzi _{1,

2}

Affiliation

The structural organization of the Golgi stacks in mammalian cells is intrinsically linked to function, including glycosylation, but the role of morphology is less clear in lower eukaryotes. Here we investigated the link between the structural organization of the Golgi and secretory pathway function using Pichia pastoris as a model system. To unstack the Golgi cisternae, we disrupted 18 genes encoding proteins in the secretory pathway without loss of viability. Using biosensors, confocal microscopy and transmission electron microscopy we identified three strains with irreversible perturbations in the stacking of the Golgi cisternae, all of which had disruption in genes that encode proteins with annotated function as or homology to calcium/calcium permeable ion channels. Despite this, no variation in the secretory pathway for ER size, whole cell glycomics or recombinant protein glycans was observed. Our investigations showed the robust nature of the secretory pathway in P. pastoris and suggest that Ca²⁺ concentration, homeostasis or signalling may play a significant role for Golgi stacking in this organism and should be investigated in other organisms.

中文翻译：

RSN1，TVP18或CSC1-2的敲除会导致巴斯德毕赤酵母中的高尔基水箱扰动

高尔基体在哺乳动物细胞中的结构组织与功能（包括糖基化）内在相关，但是在低等真核生物中，形态学的作用尚不清楚。在这里，我们调查了使用毕赤酵母的高尔基体的结构组织和分泌途径功能之间的联系作为模型系统。为了拆开高尔基水箱，我们破坏了分泌途径中编码蛋白质的18个基因，而没有丧失生存能力。使用生物传感器，共聚焦显微镜和透射电子显微镜，我们鉴定出了三个在高尔基池中堆叠时具有不可逆扰动的菌株，所有这些菌株在编码具有注释功能的蛋白质的基因中都有破坏，这些蛋白质具有与钙/钙可渗透离子通道相同的功能或同源性。尽管如此，在ER大小，全细胞糖组学或重组蛋白聚糖的分泌途径中未观察到变化。我们的研究显示了巴斯德毕赤酵母分泌途径的强大特性，并表明Ca ²⁺ 浓度，体内稳态或信号传导可能在该生物中的高尔基体堆积中起重要作用，应在其他生物中进行研究。

更新日期：2020-12-02

点击分享查看原文

点击收藏

阅读更多本刊最新论文本刊介绍/投稿指南11