权益分类	功能权益	普通用户	{{item.name}}会员
{{category.name}}	{{benefitItem.name}}

IDENTIFICATION OF NUCLEAR PATHWAYS THAT ARE CAUSALLY INVOLVED IN NUCLEOLAR TARGETING OF NF-KAPPAB/RELA.

鉴定与 NF-KAPPAB/RELA 核仁靶向相关的核通路。

基本信息

批准号：
MR/J001481/1
负责人：
Lesley Stark
金额：
$ 42.58万
依托单位：
University of Edinburgh
依托单位国家：
英国
项目类别：
Research Grant
财政年份：
2012
资助国家：
英国
起止时间：
2012 至无数据
项目状态：
已结题

来源：
https://gtr.ukri.org/projects?ref=MR%2FJ001481%2F1
关键词：
IDENTIFICATION NUCLEAR PATHWAYS CAUSALLY INVOLVED

项目摘要

Colorectal cancer is the most common cause of cancer death amongst the non-smoking population in the UK and is a major public health issue for the British population. Unequivocal evidence indicates that aspirin and related agents can prevent colorectal cancer and cause regression of this disease. However, the potential of aspirin-like agents is limited by their toxicity. In this lab, we have been undertaking studies to understand how aspirin-like agents act against colon cancer cells, in order to develop more effective and specific alternatives. We have shown that these agents cause a molecule involved in the regulation of cell growth and death, RelA, to move from the cytoplasmic compartment of the cell to the nucleoplasmic compartment then to a nuclear compartment called the nucleolus. We have also shown that movement of RelA from the nucleoplasm to the nucleolus is important for the ability of aspirin-like agents to kill colon cancer cells. Furthermore, we have demonstrated that artificially localising RelA to the nucleolus mediates the death of this cell typeThe overall objective of this study is to understand how aspirin causes RelA to move from the nucleoplasm to the nucleolus. Identifying the cellular pathways involved will allow the design of preventative/therapeutic agents that force RelA into the nucleolar compartment to kill colon cancer cells. We have already shown that a molecule called COMMD1 is important in causing RelA to go to the nucleolus in response to aspirin and that COMMD1 acts by linking a regulatory molecule called ubiquitin, to RelA.The specific objectives of this proposal are:1. To identify molecules that, alongside COMMD1, are important for linking ubiquitin to RelA after aspirin treatment. We will examine the role of candidate molecules that have previously been identified as playing a role in linking ubiquitin to RelA. We will also isolate COMMD1 from aspirin treated cells and use a technique called mass spectrometry to identify molecules that bind to COMMD1 in response to the agent. We will examine how aspirin effects these molecules and how aspirin upregulates COMMD1.2. Identification of the specific parts of RelA that are linked to ubiquitin in response to aspirin and the nature of this linkage. Ubiquitin is generally linked onto proteins at specific sites called lysines. We will identify the lysines of RelA that ubiquitin is linked to in response to aspirin. Ubiquitin also links to itself on lysine molecules to form chains. We will determine which lysine on ubiquitin is critical for nucleolar translocation of RelA using, amongst other approaches, a single cell assay where ubiquitination can be visualised.3. Identification of proteins that transport ubiquitin-linked RelA to the nucleolus. When ubiquitin is linked to a molecule, specific proteins bind to that molecule. Therefore, we suggest that when ubiquitin is linked to RelA, specific molecules bind RelA and transport the protein to the nucleolus. We will use a labelling approach and mass spectrometry to identify proteins that bind specifically to aspirin-induced, ubiquitin-linked RelA. These complementary approaches will allow us to understand the basic science of how RelA is regulated in the nucleoplasm, how linking ubiquitin to RelA causes it to go to the nucleolus and how proteins similar to RelA locate to different compartments in the cell. More importantly, these studies may reveal a way to chemically manipulate these pathways to mimic effects of aspirin on colon cancer cells.

结直肠癌是英国非吸烟人群中最常见的癌症死亡原因，也是英国人口的主要公共卫生问题。明确的证据表明，阿司匹林和相关药物可以预防结直肠癌，并导致这种疾病的消退。然而，阿司匹林样药物的潜力受到其毒性的限制。在这个实验室中，我们一直在进行研究，以了解阿司匹林样药物如何对抗结肠癌细胞，以开发更有效和更具体的替代品。我们已经表明，这些代理商导致参与细胞生长和死亡的调节，RelA，从细胞的细胞质室移动到核质室，然后到称为核仁的核室的分子。我们还表明，RelA从核质到核仁的运动对于阿司匹林样药物杀死结肠癌细胞的能力是重要的。此外，我们已经证明，人工本地化RelA的核仁介导的死亡，这种细胞typeThe本研究的总体目标是了解阿司匹林如何导致RelA从核质移动到核仁。确定所涉及的细胞途径将允许设计预防/治疗剂，迫使RelA进入核仁区室以杀死结肠癌细胞。我们已经表明，一种名为COMMD 1的分子在引起RelA响应阿司匹林而进入核仁中是重要的，COMMD 1通过连接一种名为泛素的调节分子与RelA起作用。确定与COMMD 1一起，在阿司匹林治疗后将泛素连接到RelA的重要分子。我们将研究候选分子的作用，以前已被确定为发挥作用，连接泛素到RelA。我们还将从阿司匹林处理的细胞中分离COMMD 1，并使用一种称为质谱的技术来识别与COMMD 1结合的分子。我们将研究阿司匹林如何影响这些分子以及阿司匹林如何上调COMMD1.2。确定响应阿司匹林与泛素连接的RelA的特定部分以及这种连接的性质。泛素通常在称为赖氨酸的特定位点连接到蛋白质上。我们将确定RelA的赖氨酸，泛素是连接到响应阿司匹林。泛素也连接到赖氨酸分子上形成链。我们将确定泛素上的赖氨酸对RelA的核仁易位至关重要，使用单细胞测定法，其中可以观察到泛素化。将泛素连接的RelA转运到核仁的蛋白质的鉴定。当泛素与一个分子连接时，特定的蛋白质与该分子结合。因此，我们认为，当泛素连接到RelA，特定的分子结合RelA和运输蛋白质的核仁。我们将使用标记方法和质谱法来鉴定与阿司匹林诱导的、泛素连接的RelA特异性结合的蛋白质。这些互补的方法将使我们能够了解RelA如何在核质中调节的基础科学，将泛素连接到RelA如何使其进入核仁，以及与RelA相似的蛋白质如何定位到细胞中的不同隔室。更重要的是，这些研究可能揭示了一种化学操纵这些途径的方法，以模拟阿司匹林对结肠癌细胞的作用。