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.

项目摘要 在性繁殖生物中，稳定基因组的遗传依赖于减数分裂，这是一个专业的细胞 从二倍体细胞产生单倍配子的分裂。在此过程中的缺陷导致异常染色体 数字，也称为非整倍性，这是不育和发育障碍的主要原因 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对，突触， 并形成跨界。突触的过程是由称为蛋白质结构的形成来定义的 Synaptonemal复合物（SC），将两个同源物连接在一起，并用作交叉的脚手架 重组。 SC由两个平行的染色质相关轴向元素和一个中央 区域由横向元素组成，这些元素以拉链般的方式连接同源物。尽管它具有结构性 在大多数真核生物中的保护，对管理SC组装及其的机制知之甚少 功能。该建议的目的是使用线虫C确定SC的结构和功能。 秀丽隐杆菌作为模型生物。秀丽隐杆线虫中的横向元素至少由四个盘绕螺旋圈组成 蛋白质，SYP-1，SYP-2，SYP-3和SYP-4，它们相互依存于它们的组装。我最近有 发现了SC，SYP-5和SYP-6的两个新组件，它们彼此寄生并播放 突触中的冗余角色。在AIM 1中，我将扩展这些发现并确定高度的重要性 SYP-5/6的保守无序c-末端。秀丽隐杆线虫的最新证据表明SC具有液体状 性质，从而使远程信号转导能够介导整个染色体的跨界控制。我 将检查SYP-5/6的C末端尾是否有助于SC和交叉的这种动态行为 规定。在AIM 2中，我将使用SYP蛋白配合物确定SC的生化特性 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组件之间的蛋白质相互作用，并确定其形成的倾向和要求 聚合物或液体状液滴。总体而言，这项工作将为SC的基本原则提供关键的见解 组织及其功能将在包括人类在内的物种中广泛应用。通过工作 在此应用中提出的建议，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，出色的培训教师和职业发展 计划，约翰·霍普金斯大学（Johns Hopkins University）是将我发展成独立科学家的理想场所。