Nuclear matrix support of DNA replication and transcription

背景自从光学显微镜被允许以来，细胞核结构的可视化一直是癌症诊断的主要方法。然而，关于引起异常核结构的潜在分子变化，或者这些变化对核过程组织的影响，人们知之甚少。核组织中固有的表观遗传信息也可能与我们理解衰老过程中的核衰变以及衰老的开始有关。目的一些参与DNA复制调控的蛋白质在正常细胞中通过附着在核基质上而在癌细胞(或未分化细胞)中固定。这个项目将询问调节转录的蛋白质是否也是如此，以测试已发表的观察结果可以在多大程度上推广到DNA复制之外。这将是重要的，以进一步测试分化过程的一部分是固定在核空间的DNA新陈代谢过程，以使模式形成的想法。它还将详细研究CIZ1蛋白，以确定它通常与核基质中的蛋白质结合。与细胞骨架不同，目前对于核基质的主要蛋白质成分还没有达成共识。新奇性和及时性的CIZ1与几种常见的人类癌症类型以及慢性年龄相关疾病有关，并正在被开发为癌症血液测试的基础。它的互动伙伴可能会提供类似的机会，作为核完整性的标志(用于再生医学策略或癌细胞分析)，并可能为治疗药物靶标提供新的机会。很少有分子可用于与核基质功能相关的研究，这使得这项工作既新颖又及时。该项目将哺乳动物细胞生物学与分析生物化学相结合，并在DNA复制和转录这两个核心过程中将主管与相关但互补的专业知识结合在一起。广泛的证据表明，DNA复制的组织与模板的使用有着内在的联系，这使得这种合作可能会对核基质的机械相关性产生新的见解。该项目将涉及哺乳动物细胞培养和同步技术、亚细胞分离和稳定性分析，使用一系列技术、荧光显微镜和蛋白质相互作用研究，包括识别核基质的新成分的2D-LC-MS。这对学生来说是一个跨越细胞生物学/生物化学界面进行潜在高影响项目的绝佳机会。学生将酌情参加两个实验的实验室会议，因此将获得向具有不同专业知识/兴趣的观众展示他们的工作的经验。两个小组还将举行联合实验室会议，这可能会引发进一步的合作和机会。这种方法正在成为常态，早期职业科学家在这种新的工作方式中获得经验是至关重要的。

Background Visualization of the structure of nuclei has been the main method of cancer diagnosis since light microscopy allowed. However, very little is known about the underlying molecular alterations that give rise to aberrant nuclear architecture, or the effect these have on the organisation of nuclear processes. The epigenetic information inherent in nuclear organisation is also likely to be relevant to our understanding of nuclear decay during aging, and the onset of senescence.Objectives Some proteins that are involved in the regulation of DNA replication are immobilized by attachment to the nuclear matrix in normal cells, but not in cancer cells (or in undifferentiated cells). This project will ask whether the same is true for proteins that regulate transcription, in order to test the extent to which published observations can be generalized beyond DNA replication. This will be important to test further the idea that part of the process of differentiation is to fix processes of DNA metabolism in nuclear space to enable pattern formation.It will also look in detail at the CIZ1 protein to identify the proteins that it normally binds to in the nuclear matrix. Unlike the cytoskeleton, there is currently no consensus on the main protein components of the nuclear matrix.Novelty and Timeliness CIZ1 has been implicated in several common human cancer types, as well as chronic age-related disorders, and is being developed as the basis for a blood test for cancer. Its interaction partners are likely to offer similar opportunities for exploitation either as markers of nuclear integrity (for application in regenerative medicine strategies or analysis of cancer cells), and might offer new opportunities for therapeutic drug targets. There are very few molecules that can be used for function-related studies of the nuclear matrix making this work both novel and timely.The project combines mammalian cell biology with analytical biochemistry, and combines supervisors with related but complementary expertise in the two core process of DNA replication and transcription. Extensive evidence shows that the organization of DNA replication is intrinsically linked to template usage, making this collaboration one that could yield new insight into the mechanistic relevance of the nuclear matrix. The project will involve mammalian cell culture and synchrony techniques, sub-cellular fractionation and stability assays to isolate nuclear matrix fractions using a range of techniques, fluorescence microscopy and protein interaction studies, including 2D-LC-MS to identify new components of the nuclear matrix.This is an excellent opportunity for a student to work across the cell biology/biochemistry interface on a potentially high impact project. The student will attend lab meetings in both labs as appropriate, and will therefore gain experience in presenting their work to audiences with different expertise/interests. There will also be joint lab meetings between the two groups, which are likely to trigger further collaborations and opportunities. Such approaches are becoming the norm and it is essential that early career scientists gain experience in this new way of working.