Investigating the Molecular Architecture of the Synaptonemal Complex and its Role in Crossover Formation.

研究联会复合体的分子结构及其在交叉形成中的作用。

• 批准号: 10220868
• 负责人: Matthew Hurlock
• 金额: $ 4.49万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-08-01 至 2022-07-14
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10220868
• 关键词: Address; Affinity Chromatography; Aneuploidy; Animal Model; Appearance; Architecture; Behavior; Biochemical; C-terminal; Caenorhabditis elegans; Cell division; Cells; Cellular biology; Characteristics; Chromatin; Chromosome Pairing; Chromosome Segregation; Chromosomes; Coiled-Coil Domain; Defect; Development; Diploid Cells; Disease; Down Syndrome; Elements; Ensure; Environment; Eukaryota; Failure; Fostering; Generations; Genetic; Genetic Crossing Over; Genetic Recombination; Genetic Variation; Genome; Germ Cells; Goals; Haploidy; Higher Order Chromatin Structure; Homologous Gene; Human; Infertility; Lead; Learning; Link; Liquid substance; Mediating; Meiosis; Molecular; Nematoda; Organism; Phosphorylation Site; Play; Polymers; Positioning Attribute; Process; Production; Program Development; Property; Prophase; Protein Biochemistry; Protein C; Proteins; Recombinant Proteins; Regulation; Reproductive Biology; Research; Role; Scientist; Signal Transduction; Spontaneous abortion; Stretching; Structure; Synapses; Synaptonemal Complex; Syp protein; Tail; Teacher Professional Development; Tertiary Protein Structure; Universities; Work; career development; chromosome number abnormality; developmental disease; genetic information; insight; novel; protein complex; protein protein interaction; protein structure; scaffold; skills; stoichiometry; transmission process

Project Summary In sexually reproducing organisms, inheritance of a stable genome relies on meiosis, a specialized cell division that produces haploid gametes from a diploid cell. Defects in this process lead to abnormal chromosome numbers, also known as aneuploidy, and this is a major cause of infertility and developmental disorders such as Down Syndrome. Successful segregation of chromosomes during meiosis requires that homologs pair, synapse, and form crossovers. The process of synapsis is defined by the formation of a proteinaceous structure called the synaptonemal complex (SC), which links two homologs together and serves as a scaffold for crossover recombination. The SC consists of two parallel stretches of chromatin-associated axial elements and a central region comprised of transverse elements, which connect homologs in a zipper-like fashion. Despite its structural conservation across most eukaryotes, little is known about the mechanisms governing SC assembly and its functions. The goal of this proposal is to determine the structure and functions of the SC using the nematode C. elegans as a model organism. The transverse elements in C. elegans are comprised of at least four coiled-coil proteins, SYP-1, SYP-2, SYP-3, and SYP-4, which are interdependent for their assembly. I have recently discovered two new components of the SC, SYP-5 and SYP-6, which are paralogous to each other and play redundant roles in synapsis. In Aim 1, I will extend these findings and determine the significance of highly conserved disordered C-termini of SYP-5/6. Recent evidence in C. elegans suggests that the SC has liquid-like properties, thereby enabling long-range signal transduction to mediate a chromosome-wide crossover control. I will examine whether the C-terminal tails of SYP-5/6 contribute to this dynamic behavior of the SC and crossover regulation. In Aim 2, I will determine the biochemical characteristics of the SC using the SYP protein complexes purified from C. elegans as well as the recombinant proteins. I will determine the stoichiometry and protein- protein interactions among the SC components and determine their propensity and requirements to form polymers or liquid-like droplets. Overall, this work will provide key insights into the fundamental principles of SC organization and its functions, which will be broadly applied across species, including humans. Through the work proposed in this application, I will learn key experimental skills in genetics, cell biology, and protein biochemistry. Combined with the state-of-the-art research environment, excellent training faculty, and career development programs, Johns Hopkins University is an ideal place to foster my development into an independent scientist.

项目概要 在有性生殖生物体中，稳定基因组的遗传依赖于减数分裂，这是一种特殊的细胞 从二倍体细胞产生单倍体配子的分裂。这个过程的缺陷会导致染色体异常 数字，也称为非整倍体，这是不孕和发育障碍的主要原因，例如 唐氏综合症。减数分裂过程中染色体的成功分离需要同源物配对、突触、 并形成交叉。突触的过程是由称为“突触”的蛋白质结构的形成来定义的。 联会复合体 (SC)，将两个同源物连接在一起并充当交叉的支架 重组。 SC 由两条平行的染色质相关轴向元件和一个中央部分组成 由横向元件组成的区域，横向元件以拉链状方式连接同系物。尽管其结构 大多数真核生物的保护性，人们对 SC 组装及其控制机制知之甚少。 功能。该提案的目标是利用线虫 C 确定 SC 的结构和功能。 线虫作为模式生物。秀丽隐杆线虫的横向元件由至少四个螺旋线圈组成 蛋白质 SYP-1、SYP-2、SYP-3 和 SYP-4，它们的组装是相互依赖的。我最近有 发现了 SC 的两个新组件 SYP-5 和 SYP-6，它们彼此是旁系同源的并发挥作用 突触中的冗余角色。在目标 1 中，我将扩展这些发现并确定高度的重要性 SYP-5/6 保守的无序 C 末端。最近在秀丽隐杆线虫中的证据表明 SC 具有液体状 特性，从而使远程信号转导能够介导染色体范围的交叉控制。我 将检查 SYP-5/6 的 C 末端尾部是否有助于 SC 和交叉的动态行为 规定。在目标 2 中，我将使用 SYP 蛋白复合物确定 SC 的生化特征 从秀丽隐杆线虫以及重组蛋白中纯化。我将确定化学计量和蛋白质- SC 成分之间的蛋白质相互作用，并确定它们形成的倾向和要求 聚合物或液体状液滴。总的来说，这项工作将为 SC 的基本原理提供重要的见解 组织及其职能，将广泛应用于包括人类在内的各个物种。通过工作 在这个应用程序中，我将学习遗传学、细胞生物学和蛋白质生物化学方面的关键实验技能。 结合最先进的研究环境、优秀的培训师资队伍和职业发展 约翰·霍普金斯大学是培养我成为一名独立科学家的理想场所。