Subcellular Localization of Matrin 3 Containing Mutations Associated with ALS and Distal Myopathy
Subcellular Localization of Matrin 3 Containing Mutations Associated with ALS and Distal Myopathy
复制标题
DOI：
10.1371/journal.pone.0142144
复制
发表时间：
2015-11-03
期刊：
影响因子：
3.7
通讯作者：
Borchelt, David R.
中科院分区：
文献类型：
--
作者：
Gallego-Iradi, M. Carolina;Clare, Alexis M.;Borchelt, David R.
背景Matrin 3 [MATR 3]是一种RNA和DNA结合蛋白，通常定位于细胞核，其突变与肌萎缩侧索硬化症（ALS）和远端肌病有关。在本研究中，我们已经使用了瞬时转染培养的细胞系来研究不同的致病突变的影响，对本地化的Matrin 3内cells.ResultsUsing CHO和人类H4神经胶质瘤细胞模型，我们发现，ALS/肌病突变不产生深刻的变化，在本地化的蛋白质。尽管我们通过免疫染色或荧光标记蛋白的可视化观察到细胞质中Matrin 3的可变水平，但大多数表达野生型（WT）或突变型Matrin 3的细胞显示该蛋白的核定位。当在细胞质中观察到细胞质免疫染色或融合蛋白荧光时，通常在细胞核中可见较强的染色或荧光强度。在类似于80%的细胞用亚砷酸钠（Ars）诱导细胞质应激颗粒，WT和F115 C突变体Matrin 3的核定位不受干扰。值得注意的是，过度表达的突变体Matrin 3并没有诱导形成明显的大包容性的结构，无论是在细胞质或nucleus.ConclusionsOur的研究结果表明，在Matrin 3的突变与ALS和肌病不显着改变蛋白质的正常定位或容易诱导包容性的形成。
BackgroundMutations in Matrin 3 [MATR3], an RNA-and DNA-binding protein normally localized to the nucleus, have been linked to amyotrophic lateral sclerosis (ALS) and distal myopathies. In the present study, we have used transient transfection of cultured cell lines to examine the impact of different disease-causing mutations on the localization of Matrin 3 within cells.ResultsUsing CHO and human H4 neuroglioma cell models, we find that ALS/myopathy mutations do not produce profound changes in the localization of the protein. Although we did observe variable levels of Matrin 3 in the cytoplasm either by immunostaining or visualization of fluorescently-tagged protein, the majority of cells expressing either wild-type (WT) or mutant Matrin 3 showed nuclear localization of the protein. When cytoplasmic immunostaining, or fusion protein fluorescence, was seen in the cytoplasm, the stronger intensity of staining or fluorescence was usually evident in the nucleus. In similar to 80% of cells treated with sodium arsenite (Ars) to induce cytoplasmic stress granules, the nuclear localization of WT and F115C mutant Matrin 3 was not disturbed. Notably, over-expression of mutant Matrin 3 did not induce the formation of obvious large inclusion-like structures in either the cytoplasm or nucleus.ConclusionsOur findings indicate that mutations in Matrin 3 that are associated with ALS and myopathy do not dramatically alter the normal localization of the protein or readily induce inclusion formation.