A Missense Mutation in the NSF Gene Causes Abnormal Golgi Morphology in Arabidopsis thaliana.
A Missense Mutation in the NSF Gene Causes Abnormal Golgi Morphology in Arabidopsis thaliana.
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DOI：
10.1247/csf.17026
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发表时间：
2018-02
影响因子：
1.5
通讯作者：
Sayuri Tanabashi;Keiko Shoda;C. Saito;T. Sakamoto;T. Kurata;T. Uemura;A. Nakano
中科院分区：
文献类型：
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作者：
Sayuri Tanabashi;Keiko Shoda;C. Saito;T. Sakamoto;T. Kurata;T. Uemura;A. Nakano
高尔基体是糖基化和膜运输的关键站。在大多数真核生物中，它由堆叠的池子组成。然而，高尔基体堆叠是如何形成和维持的机制仍然不清楚。模式植物拟南芥提供了一个很好的系统来用光学显微镜观察高尔基体结构，因为拟南芥中的高尔基体呈迷你堆叠的形式分布在细胞质中。为了获得了解高尔基体形态的分子基础的线索，我们采取了正向遗传的方法来分离在共聚焦显微镜下显示高尔基体异常结构的突变体。在本报告中，我们描述了一个这样的突变体的特征，命名为#46-3。46-3号突变体表现出多效性高尔基体表型。高尔基体的大小大多小于野生型，但在共聚焦显微镜下从非常小的，有时没有明显的顺式和反式脑池联系到异常大的。在超微结构水平上，偶尔可以观察到奇怪的大高尔基体堆。通过位置定位、基因组测序、互补和等位基因测试，我们将突变表型与NSF基因中的错义突变D374N联系起来，该错义突变编码N-乙基马来酰亚胺敏感因子(NSF)，是膜融合的关键成分。该残基位于NSF的ATP结合位点附近，NSF在真核生物中非常保守，这表明NSF的生化功能对于维持高尔基体的正常形态非常重要。关键词：高尔基体形态，N-乙基马来酰亚胺敏感因子(NSF)，拟南芥。
The Golgi apparatus is a key station of glycosylation and membrane traffic. It consists of stacked cisternae in most eukaryotes. However, the mechanisms how the Golgi stacks are formed and maintained are still obscure. The model plant Arabidopsis thaliana provides a nice system to observe Golgi structures by light microscopy, because the Golgi in A. thaliana is in the form of mini-stacks that are distributed throughout the cytoplasm. To obtain a clue to understand the molecular basis of Golgi morphology, we took a forward-genetic approach to isolate A. thaliana mutants that show abnormal structures of the Golgi under a confocal microscope. In the present report, we describe characterization of one of such mutants, named #46-3. The #46-3 mutant showed pleiotropic Golgi phenotypes. The Golgi size was in majority smaller than the wild type, but varied from very small ones, sometimes without clear association of cis and trans cisternae, to abnormally large ones under a confocal microscope. At the ultrastructual level by electron microscopy, queer-shaped large Golgi stacks were occasionally observed. By positional mapping, genome sequencing, and complementation and allelism tests, we linked the mutant phenotype to the missense mutation D374N in the NSF gene, encoding the N-ethylmaleimide-sensitive factor (NSF), a key component of membrane fusion. This residue is near the ATP-binding site of NSF, which is very well conserved in eukaryotes, suggesting that the biochemical function of NSF is important for maintaining the normal morphology of the Golgi.Key words: Golgi morphology, N-ethylmaleimide-sensitive factor (NSF), Arabidopsis thaliana.