Formation of a nuclear complex containing the p53 tumor suppressor, YB-1, and the Werner syndrome gene product in cells treated with UV light
Formation of a nuclear complex containing the p53 tumor suppressor, YB-1, and the Werner syndrome gene product in cells treated with UV light
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DOI：
10.1016/j.biocel.2006.01.008
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发表时间：
2006-01-01
影响因子：
4
通讯作者：
Lebel, Michel
中科院分区：
文献类型：
--
作者：
Guay, David;Gaudreault, Isabelle;Lebel, Michel
YB-1是一种多功能蛋白，参与转录、翻译和mRNA剪接的调节。近年来，一些实验室已经证明YB-1也直接参与了细胞对遗传毒性应激的反应。因此，一份报告表明，沃纳综合征基因产物(WRN)是从YB-1亲和层析柱中洗脱出来的。沃纳综合征是一种罕见的疾病，其特征是过早发作一些与年龄相关的疾病，包括癌症。导致Werner综合征的基因编码一种DNA解旋酶/核酸外切酶蛋白，据信参与了与P53的DNA修复的某些方面。在这项研究中，我们证明了肿瘤抑制因子P53在体外连接了WRN和YB-1蛋白。荧光标记蛋白质的显微镜分析和免疫共沉淀实验证实，在人类细胞中形成了YB-1/P53/WRN复合体，但只有在紫外光处理后才能形成。我们还证实了在紫外线照射下，P53是GFP-YB-1融合蛋白从细胞质转移到含有WRN蛋白的几个核焦点的主要参与者。在用拓扑异构酶抑制剂依托泊苷或拟放射药物博莱霉素处理的细胞中，这种易位没有发生。这些结果表明，YB-1/P53/WRN复合体是对细胞中特定DNA损伤的反应而形成的。(C)2006爱思唯尔有限公司。保留所有权利。
YB-1 is a multifunctional protein involved in the regulation of transcription, translation, and mRNA splicing. In recent years, several laboratories have demonstrated that YB-1 is also directly involved in the cellular response to genotoxic stress. Accordingly, one report has indicated that the Werner syndrome gene product (WRN) is eluted from an YB-1 affinity chromatography column. Werner syndrome is a rare disorder characterized by the premature onset of a number of age-related diseases, including cancer. The gene responsible for Werner syndrome encodes a DNA helicase/exonuclease protein believed to be involved in some aspect of DNA repair with p53. In this study, we demonstrate that the tumor suppressor, p53, bridges the WRN and YB-1 proteins in vitro. Microscopic analyses of fluorescent-tagged proteins and co-immunoprecipitation experiments confirmed the formation of an YB-1/p53/WRN complex in human cells, but only after treatment with UV light. We also confirmed that p53 is a major player in the translocation of GFP-YB-1 fusion proteins from the cytoplasm to several nuclear foci containing WRN proteins upon UV irradiation. Such translocation did not occur in cells treated with the topoisomerase inhibitor, etoposide, or the radiomimetic drug, bleomycin. Such results suggest that an YB-1/p53/WRN complex is formed in response to the emergence of specific DNA lesions in cells. (c) 2006 Elsevier Ltd. All rights reserved.