Determinants of Centromere Identity in Drosophila

描述(申请人提供)：染色体复制和传递对于遗传特性是必不可少的，但在多细胞真核生物中，负责这些过程的机制仍然知之甚少。着丝粒是有丝分裂和减数分裂过程中着丝粒的关键附着部位，在中期染色体上表现为一种皱缩现象。现在着丝粒领域的一个紧迫问题是着丝粒身份是如何在多细胞真核生物中从一代传播到下一代的。要阐明着丝粒识别、繁殖和功能的决定因素，需要鉴定促进着丝粒形成和反式功能的基因产物，并最终确定它们如何与顺式作用元件相互作用来发挥其基本功能。本质上，我们需要了解着丝粒染色质的组织、复制和修饰。 在高等真核生物中解决这些复杂问题所需的多方面的遗传、分子、细胞生物学和生化方法很可能在果蝇身上取得成功。使用果蝇进行实验分析已经有一个多世纪的历史，这有助于进行复杂的活体分析。在这里，我们提出了遗传、分子、细胞生物学和生化实验，旨在鉴定和鉴定促进着丝粒染色质组装和繁殖的基因产物，并确定它们的性质和功能。我们进入着丝粒染色质的切入点将是一种保守的组蛋白H3样蛋白(CID，着丝粒识别符)，它仅定位于功能着丝粒。我们将使用遗传筛选和生化分离来鉴定与CID相互作用的基因和蛋白质，然后使用各种方法来确定它们在着丝粒功能中的作用。这些研究将解决特定的假设，并为未来分析果蝇和其他高等真核生物(如人类)的遗传和着丝粒功能奠定基础。

DESCRIPTION (provided by applicant): Chromosome replication and transmission are essential for the inheritance of genetic traits, but the mechanisms responsible for these processes remain poorly understood in multicellular eukaryotes. The centromere, which appears as a constriction in metaphase chromosomes, is associated with the kinetochore and serves as the key attachment site to the spindle during mitosis and meiosis. A pressing question in the centromere field today is how centromere identity is propagated from one generation to the next in multicellular eukaryotes. Elucidating the determinants of centromere identity, propagation and function requires identification of the gene products that promote centromere formation and function in trans, and ultimately determining how they interact with the cis-acting elements to perform their essential functions. In essence, we need to understand the organization, replication, and modification of centromeric chromatin. The multifaceted genetic, molecular, cell biological and biochemical approaches required to address these complex questions in higher eukaryotes are likely to succeed in Drosophila. There is over a century of experimental analyses using the fruit fly, which facilitates sophisticated in vivo analyses. Here, we propose genetic, molecular, cell biological and biochemical experiments designed to identify and characterize gene products that promote the assembly and propagation of centromeric chromatin, and to determine their properties and functions. Our entry point into centromeric chromatin will be a conserved histone H3-like protein (CID, for Centromere IDentifier) that localizes exclusively to functional centromeres. We will use genetic screens and biochemical isolations to identify genes and proteins that interact with CID, then use a variety of approaches to determine their roles in centromere function. These studies will address specific hypotheses and provide the groundwork for future analysis of inheritance and centromere function in Drosophila and other higher eukaryotes, such as humans.