Probing the mRNA processing body using protein macroarrays and "autoantigenomics".
Probing the mRNA processing body using protein macroarrays and "autoantigenomics".
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DOI：
10.1261/rna.411907
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发表时间：
2007-05
期刊：
影响因子：
4.5
通讯作者：
Wei-hong Yang;D. Bloch
中科院分区：
文献类型：
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作者：
Wei-hong Yang;D. Bloch
信使 RNA 加工体（P 体）是在 mRNA 降解中发挥直接作用的细胞结构。 P-体也与 RNAi 介导的转录后基因沉默有关。尽管 P 体在细胞生物学中发挥着重要作用，但 P 体的成分及其组织仅得到部分定义。大约 5% 的自身免疫性疾病原发性胆汁性肝硬化患者具有针对这些结构的抗体。蛋白质宏阵列技术的最新进展允许同时筛选数千种蛋白质的与自身抗体的反应性。我们使用来自具有抗 P 体自身抗体的患者的血清来筛选蛋白质宏阵列，并鉴定了 67 种潜在的自身抗原。免疫反应蛋白包括四种已知的 P 体成分和三种额外的原发性胆汁性肝硬化自身抗原。 Y-box 蛋白 1 (YB-1) 是一种 50 kDa RNA 结合蛋白，以前不知道它是 P 体成分，七名患者中有四名的血清识别出了 Y-box 蛋白 1 (YB-1)。 YB-1 与 P 体成分 DCP1a 和 Ge-1 共定位。在遭受亚砷酸盐诱导的氧化应激的细胞中，YB-1 定位于含有 TIA 的应激颗粒。在氧化应激过程中，YB-1 和先前鉴定的 P 体成分 RAP55 均从 P 体易位至应激颗粒。然而，在恢复过程中，与 RAP55 相比，YB-1 在 P 体中的重新出现延迟，表明 YB-1 和 RAP55 可能具有不同的功能。这项研究表明，人类自身抗体和蛋白质宏阵列技术的结合提供了一种鉴定和表征 mRNA P 体成分的新方法。
Messenger RNA processing bodies (P-bodies) are cellular structures that have a direct role in mRNA degradation. P-bodies have also been implicated in RNAi-mediated post-transcriptional gene silencing. Despite the important roles of P-bodies in cellular biology, the constituents of P-bodies and their organization have been only partially defined. Approximately 5% of patients with the autoimmune disease primary biliary cirrhosis have antibodies directed against these structures. Recent advances in protein macroarray technology permit the simultaneous screening of thousands of proteins for reactivity with autoantibodies. We used serum from patients with anti-P-body autoantibodies to screen a protein macroarray and identified 67 potential autoantigens. Immunoreactive proteins included four known P-body components and three additional primary biliary cirrhosis autoantigens. Y-box protein 1 (YB-1), a 50-kDa RNA-binding protein that was not previously known to be a P-body component, was recognized by serum from four of seven patients. YB-1 colocalized with P-body components DCP1a and Ge-1. In cells subjected to arsenite-induced oxidative stress, YB-1 localized to TIA-containing stress granules. Both YB-1 and the previously identified P-body component RAP55 translocated from P-bodies to stress granules during oxidative stress. During recovery, however, the reappearance of YB-1 in P-bodies was delayed compared with that of RAP55, suggesting that YB-1 and RAP55 may have different functions. This study demonstrates that the combination of human autoantibodies and protein macroarray technology provides a novel method for identifying and characterizing components of mRNA P-bodies.