USE OF YTH TO IDENTIFY INTERACTIONS WITH CENTROMERIC NUCLEOSOMES

使用 YTH 鉴定与着丝粒核小体的相互作用

• 批准号: 8171333
• 负责人: Steven Henikoff
• 金额: $ 2.21万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-09-01 至 2011-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8171333
• 关键词: Binding; Centromere; Chromatin; Computer Retrieval of Information on Scientific Projects Database; DNA; Funding; Genome; Grant; Histone H2A; Histone H4; Histones; Institution; Kinetochores; Mitosis; Modeling; Nucleosomes; Proteins; Research; Research Personnel; Resources; Solvents; Source; Structure; Surface; Testing; United States National Institutes of Health; Work; novel; particle; yeast two hybrid system

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. The eukaryotic genome is packaged into nucleosomes, particles that consist of DNA wrapped around histone proteins. Centromeres are specialized regions of chromatin that serve as points of attachment of kinetochore during mitosis. In the course of recent work it has been proposed that the structure of the centromeric histone core is fundamentally different than that of the canonical one. Canonical nucleosomes contain two molecules each of histones H2A, H2B, H3 and H4 that together comprise an octamer. However, recently obtained experimental evidence suggests that centromeric nucleosome is a smaller particle, most likely containing a single copy of histones H2A, H2B, CenH3 and H4, a hemisome. The hemisome model predicts that novel surfaces on the CenH3-containing particle that are not exposed on the canonical particle can be specifically bound by kinetochore proteins. Using the crystal structure of the canonical nucleosome as a starting point we have identified regions of H2B and CenH3 that might be exposed to solvent in a hemisome and thus be available for binding by kinetochore proteins. We would like to use the Yeast Two Hybrid system to screen for interaction partners of these proteins, using histone H4 as a control. We hope to identify proteins with known kinetochore function that will test our model and allow us to propose new hypotheses for the inner kinetochore structure.

这个子项目是许多研究子项目中的一个 由NIH/NCRR资助的中心赠款提供的资源。子项目和 研究者（PI）可能从另一个NIH来源获得了主要资金， 因此可以在其他CRISP条目中表示。所列机构为 研究中心，而研究中心不一定是研究者所在的机构。 真核基因组被包装成核小体，核小体是由DNA包裹在组蛋白周围组成的颗粒。着丝粒是染色质的特化区域，在有丝分裂期间充当动粒的附着点。在最近的工作过程中，有人提出，着丝粒组蛋白核心的结构与规范组蛋白核心的结构有根本的不同。典型的核小体含有两个组蛋白H2 A、H2 B、H3和H4分子，它们共同构成一个八聚体。然而，最近获得的实验证据表明，着丝粒核小体是一个较小的颗粒，最有可能包含组蛋白H2 A，H2 B，CenH 3和H4的单拷贝，一个半球。半球体模型预测，未暴露在典型颗粒上的含CenH 3的颗粒上的新表面可以被动粒蛋白特异性结合。使用典型核小体的晶体结构作为起点，我们已经确定了H2 B和CenH 3的区域，这些区域可能暴露于半体中的溶剂中，从而可用于结合动粒蛋白。我们希望使用酵母双杂交系统来筛选这些蛋白质的相互作用伴侣，使用组蛋白H4作为对照。我们希望确定具有已知动粒功能的蛋白质，这将测试我们的模型，并使我们能够对内部动粒结构提出新的假设。