Regulation of gene-specific nuclear NF-kappaB functions by cofactors and posttranslational modifications

辅因子和翻译后修饰对基因特异性核 NF-κB 功能的调节

• 批准号: 162103480
• 负责人: Professor Dr. Michael Kracht
• 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: 2009
• 资助国家: 德国
• 起止时间: 2008-12-31 至 2017-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/162103480?language=en
• 关键词: Regulation; gene; specific; nuclear; NF

The transcription factor NF-kappaB plays a central role for stress-induced gene regulation. The identification of the cytosolic events mediating NF-kappaB activation is rapidly progressing, whereas the gene-specific functions of this transcription factor in the nucleus are poorly understood. In our preliminary work we identified novel nuclear cofactors (CDK6 and TRIP6) and the function and regulation of several postranslational modifications of NF-kappaB p65. This preceding work forms the basis for a bipartite research program that aims to elucidate the nuclear functions of NF-kappaB in much more detail. The first part will investigate the mechanisms and signaling pathways mediating the gene regulatory functions of CDK6 and TRIP6. We will also investigate the contribution of TRIP6 phosphorylation for its function in glucocorticoid-mediated repression of NF-kappaB-dependent gene expression. Genome-wide analyses will clarify whether CDK6 is specialized to communicate cell cycle-specific signals to NF-kappaB or whether this kinase is of general relevance for a broad range of NF-kappaB activating stimuli. The second part will investigate the genome-wide recruitment of posttranslationally modified NF-kappaB p65 and a DNA-binding mutant to investigate their physiological function for transcriptional activation and chromatin association. We will also investigate the functional consequences of a stimulus-induced conformational switch of NF-kappaB p65 using a recently identified conformation-specific antibody. These experiments will allow conclusions on the significance of posttranslational modifications, context-specific conformation and DNA-binding for p65 functions. The overlapping and complementary expertise of both applicants will allow a significantly improved understanding of the molecular mechanisms underlying the plasticity of NF-kappaB-dependent gene regulation in healthy and diseased cells.

转录因子NF-κ B在胁迫诱导的基因调控中起着重要作用。鉴定介导NF-κ B活化的胞质事件正在迅速进展，而这种转录因子在细胞核中的基因特异性功能知之甚少。在我们的初步工作中，我们鉴定了新型核辅因子（CDK 6和TRIP 6）以及NF-kappaB p65的几种翻译后修饰的功能和调节。这项工作为一项旨在更详细地阐明NF-κ B核功能的双边研究计划奠定了基础。第一部分研究CDK 6和TRIP 6的基因调控机制和信号通路。我们还将研究TRIP 6磷酸化在糖皮质激素介导的NF-κ B依赖性基因表达抑制中的作用。全基因组分析将阐明CDK 6是否专门将细胞周期特异性信号传递给NF-κ B，或者这种激酶是否与广泛的NF-κ B激活刺激物具有普遍相关性。第二部分将研究全基因组范围内的招聘后修饰的NF-κ B p65和DNA结合突变体，以研究其生理功能的转录激活和染色质协会。我们还将调查的刺激诱导的NF-κ B p65的构象转换的功能后果，使用最近确定的构象特异性抗体。这些实验将允许的翻译后修饰，上下文特异性构象和DNA结合的p65功能的意义上的结论。这两个申请人的重叠和互补的专业知识将允许显着提高对健康和患病细胞中NF-κ B依赖性基因调控可塑性的分子机制的理解。