Genetic and Epigenetic Determinants of Centromere Identity

着丝粒身份的遗传和表观遗传决定因素

• 批准号: 10631092
• 负责人: BARBARA MELLONE
• 金额: $ 40.25万
• 依托单位: UNIVERSITY OF CONNECTICUT STORRS
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-08-01 至 2025-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10631092
• 关键词: Animals; Biological Models; Cell division; Cells; Centromere; Chromatin; Chromosome Segregation; Chromosome Structures; Chromosomes; DNA; Defect; Development; Drosophila genus; Elements; Engineering; Epigenetic Process; Fertility; Genetic; Goals; Health; Histones; Human; Infertility; Kinetochores; Lead; Light; Maintenance; Mediating; Meiosis; Microtubules; Mitosis; Mitotic; Modeling; Nature; Organism; Process; Refractory; Role; Specific qualifier value; Spontaneous abortion; System; Testing; Variant; Work; centromere protein A; chromosome conformation capture; chromosome missegregation; daughter cell; genomic tools; tumor progression

Project Summary Centromeres are essential chromosomal elements that mediate kinetochore assembly and accurate chromosome segregation. Centromere defects lead to chromosome missegregation, with detrimental effects on cell and organism health and fertility. In most multicellular species, centromeres are composed of large regions of highly repetitive DNA marked by chromatin containing the centromere-specific histone variant CENP-A. Previous work demonstrated that both centromeric DNA and CENP-A chromatin have the potential to initiate centromere activity de novo; however, their respective contributions to centromere specification in mitosis and meiosis have remained elusive. The overall goal of this proposal is to determine how centromeric DNA and chromatin contribute to centromere identity. The centromeres of metazoans have been refractory to full sequencing and assembly due their large size and highly repetitive nature, hampering our ability to systematically interrogate the role of centromeric DNA elements. Additionally, there are currently no systems in which to test if de novo centromeres, which are devoid of centromeric DNA, and can sustain centromere function and specification through mitotic and meiotic divisions. Using our unique advancements in Drosophila, which include the identification and assembly of its centromeric sequences and the establishment of an inducible de novo centromere system, this proposal will: 1) test if chromatin-mediated centromeres can sustain chromosome segregation through development and meiosis, effectively replacing endogenous centromeres; 2) test specific hypothesis on how centromeric DNA elements may contribute to CENP-A chromatin establishment or maintenance. Collectively, this work will shed light into centromere specification mechanisms in Drosophila, an exceptional model system that allows centromere studies in the context of animal development and fertility- with broad relevance to other species, including humans.

项目摘要 着丝粒是介导动粒组装和精确定位的重要染色体元件。 染色体分离着丝粒缺陷导致染色体错误分离，具有不利影响 对细胞和有机体的健康和生育能力的影响。在大多数多细胞物种中， 由含有着丝粒特异性组蛋白变体的染色质标记的高度重复DNA区域 CENP-A。以前的工作表明，着丝粒DNA和CENP-A染色质都有潜力， 从头启动着丝粒活性;然而，它们各自对 有丝分裂和减数分裂仍然难以捉摸。这项提案的总体目标是确定着丝粒如何 DNA和染色质有助于着丝粒的同一性。后生动物的着丝粒一直难以被 由于它们的大尺寸和高度重复的性质，完全测序和组装，阻碍了我们的能力， 系统地询问着丝粒DNA元件的作用。此外，目前还没有系统 以测试没有着丝粒DNA的从头着丝粒是否可以维持着丝粒 通过有丝分裂和减数分裂的功能和规格。利用我们在果蝇方面的独特进展， 包括其着丝粒序列的鉴定和组装， 可诱导的从头着丝粒系统，该建议将：1）测试染色质介导的着丝粒是否可以维持 通过发育和减数分裂的染色体分离，有效地取代内源性着丝粒; 2） 测试关于着丝粒DNA元件如何有助于CENP-A染色质建立的特定假设 或维护。总的来说，这项工作将揭示果蝇中的着丝粒特化机制， 一个特殊的模型系统，允许在动物发育和生育能力的背景下进行着丝粒研究- 与包括人类在内的其他物种有着广泛的相关性。