Regulation of IKK-epsilon and TRAF6 functions by posttranslational modifications

通过翻译后修饰调节 IKK-epsilon 和 TRAF6 功能

• 批准号: 219706149
• 负责人: Professor Dr. M. Lienhard Schmitz
• 金额: --
• 依托单位: Biochemisches Institut
• 依托单位国家: 德国
• 项目类别: Research Grants
• 财政年份: 2012
• 资助国家: 德国
• 起止时间: 2011-12-31 至 2015-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/219706149?language=en
• 关键词: Regulation; IKK; epsilon; TRAF6; functions

Lipopolysaccaride (LPS) contained on the surface of gram-negative bacteria triggers complex intracellular signaling cascades activating transcription factors such as NF-ýB that serve to induce the expression of immune regulatory genes. The last years have witnessed the identification of many different components of the LPS-inducible intracellular signaling system, including the protein kinase IKKý and the ubiquitin E3 ligase TRAF6. As the control of their activity is not well known, this project aims to study their mutual regulation by posttranslational modifications. Preliminary results show that IKKý phosphorylates TRAF6 at 3 serines, while TRAF6 mediates inducible ubiquitination of IKKý at 9 lysines. To address the functional role of these posttranslational modifications, knock-out cells will be reconstituted to stably express these proteins in their wildtype and mutated forms that are changed in the modified amino acids. These cells will be analyzed by a large variety of experimental approaches including cell biological and biochemical experiments. The occurrence, kinetics and regulation of these modifications are traced by modification-specific antibodies. As recent evidence shows a role of IKKý in the nucleus, ChIP-Seq experiments will reveal a comprehensive map of genomic binding sites for this kinase. This basic research project will broaden our understanding of molecular processes that occur in proinflammatory signal transduction and are thus of biomedical interest.

革兰氏阴性细菌表面上包含的脂多多亚菌（LPS）会触发复杂的细胞内信号传导级联激活转录因子，例如NF-min，这些因子有助于诱导免疫调节基因的表达。最后几年见证了LPS诱导的细胞内信号传导系统的许多不同组成部分，包括蛋白激酶Ikký和泛素E3连接酶TRAF6。由于对其活动的控制尚不清楚，因此该项目旨在通过翻译后修改研究其相互调控。初步结果表明，ikký磷酸化在3丝氨酸处将TRAF6磷酸化，而Traf6在9个级别中介导了Ikký的诱导性泛素化。为了解决这些翻译后修饰的功能作用，将重组敲除细胞以稳定地以其野生型和突变形式表达这些蛋白质，这些蛋白质在改良的氨基酸中发生了变化。这些细胞将通过包括细胞生物学和生化实验在内的多种实验方法进行分析。这些修饰的发生，动力学和调节是通过修饰特异性抗体追溯的。由于最近的证据表明了Ikký在核中的作用，Chip-Seq实验将揭示该激酶的基因组结合位点的全面图。这个基础研究项目将扩大我们对促炎信号转导中发生的分子过程的理解，因此是生物医学的兴趣。

项目成果

K63-Ubiquitylation and TRAF6 Pathways Regulate Mammalian P-Body Formation and mRNA Decapping.

• DOI: 10.1016/j.molcel.2016.05.017
• 发表时间: 2016-06
• 期刊: Molecular cell
• 影响因子: 16
• 作者: U. Tenekeci;M. Poppe;K. Beuerlein;C. Buro;H. Müller;Hendrik Weiser;D. Kettner-Buhrow;Katharina Porada;Doris Newel;Ming Xu;Zhijian J. Chen-Zhijian J.-Chen-2248993093;J. Busch;M. L. Schmitz;M. Kracht