Comparing lamin proteins post-translational relative stability using a 2A peptide-based system reveals elevated resistance of progerin to cellular degradation.
Comparing lamin proteins post-translational relative stability using a 2A peptide-based system reveals elevated resistance of progerin to cellular degradation.
复制标题
使用基于 2A 肽的系统比较核纤层蛋白翻译后的相对稳定性，揭示了早老素对细胞降解的抵抗力增强。
DOI：
10.1080/19491034.2016.1260803
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发表时间：
2016
期刊：
影响因子：
--
通讯作者：
Cao,Kan
中科院分区：
文献类型：
--
作者：
Wu,Di;Yates,PhillipA;Zhang,Haoyue;Cao,Kan
核纤层蛋白是细胞核外围核纤层的主要组成部分，支持核膜并参与许多核过程，包括 DNA 复制、转录和染色质组织。核纤层蛋白病这一类疾病与核纤层蛋白基因突变有关。最引人注目的病例之一是哈钦森-吉尔福德早衰综合症 (HGPS)，这是一种名为早衰蛋白的核纤层蛋白 A 显性失活突变体的结果。由于永久性 C 末端法呢基尾部的异常存在，早老素逐渐在核膜上积聚，扰乱多种信号传导和转录事件。早老蛋白的积累导致人们推测早老蛋白具有比野生型核纤层蛋白A蛋白更高的稳定性。然而，核纤层蛋白的低溶解度使得传统的免疫沉淀依赖性方法（例如脉冲追踪分析）无法有效比较突变型和野生型核纤层蛋白的相对稳定性。在这里，我们采用了一个新的平台来推断核纤层蛋白稳定性的差异，该平台基于猪 teschovirus-1 2A 肽介导的共翻译切割后的共翻译报告蛋白的标准化。使用该方法获得的结果支持早老蛋白比核纤层蛋白 A 更稳定的观点。此外，FTI 处理将早老蛋白的相对稳定性降低至野生型核纤层蛋白 A 的水平。
Nuclear lamins are the major components of the nuclear lamina at the periphery of the nucleus, supporting the nuclear envelope and participating in many nuclear processes, including DNA replication, transcription and chromatin organization. A group of diseases, the laminopathies, is associated with mutations in lamin genes. One of the most striking cases is Hutchinson-Gilford progeria syndrome (HGPS) which is the consequence of a lamin A dominant negative mutant named progerin. Due to the abnormal presence of a permanent C-terminal farnesyl tail, progerin gradually accumulates on the nuclear membrane, perturbing a diversity of signalings and transcriptional events. The accumulation of progerin has led to the speculation that progerin possesses higher stability than the wild type lamin A protein. However, the low solubility of lamin proteins renders traditional immunoprecipitation-dependent methods such as pulse-chase analysis ineffective for comparing the relative stabilities of mutant and wild type lamins. Here, we employ a novel platform for inferring differences in lamin stability, which is based on normalization to a co-translated reporter protein following porcine teschovirus-1 2A peptide-mediated co-translational cleavage. The results obtained using this method support the notion that progerin is more stable than lamin A. Moreover, treatment of FTI reduces progerin relative stability to the level of wild type lamin A.