Characterizing the Sexually Dimorphic Role of Topoisomerase II During the Sister Chromatid Cohesion Release Pathway.

表征拓扑异构酶 II 在姐妹染色单体凝聚释放途径中的性别二态性作用。

• 批准号: 10536795
• 负责人: Christine Kiely Rourke
• 金额: $ 4.2万
• 依托单位: UNIVERSITY OF DELAWARE
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-10-01 至 2025-09-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10536795
• 关键词: Affect; Affinity Chromatography; Alleles; Aneuploidy; Animal Model; Biochemical; Biochemistry; Caenorhabditis elegans; Cell division; Cells; Chromosome Segregation; Chromosome Structures; Chromosomes; Collaborations; Confocal Microscopy; Congenital Abnormality; Couples; DNA; Data; Delaware; Developing Countries; Diploidy; Ensure; Environment; Enzymes; Equipment; Ethnic Origin; Event; Exhibits; Faculty; Female; Female infertility; Fluorescence; Gametogenesis; Genetic; Genetic Crossing Over; Genetic Recombination; Germ Cells; Goals; Haploidy; Health; Homologous Gene; In Vitro; Infertility; Knowledge; Label; Lead; Meiosis; Meiotic Prophase I; Mentors; Mentorship; Microscopy; Mitosis; Molecular; Molecular Biology; Nationalities; Oocytes; Oogenesis; Organism; Pathway interactions; Persons; Phenotype; Physical condensation; Play; Ploidies; Postdoctoral Fellow; Prophase; Proteins; Race; Regulation; Research Personnel; Resolution; Resources; Role; Sex Differences; Sister Chromatid; Socioeconomic Status; Spermatogenesis; Spontaneous abortion; Testing; Topoisomerase II; Training; Universities; aurora B kinase; career; cohesion; egg; in vivo; innovation; insight; male; mutant; nano; novel; precursor cell; programs; protein complex; protein expression; protein protein interaction; segregation; sex; sexual dimorphism; socioeconomics; sperm cell; stoichiometry; success

PROJECT SUMMARY People in every socioeconomic class, both males and females, and every nationality and ethnicity are affected by infertility. Aneuploidy is the leading cause of infertility, as well as birth defects, and miscarriages. Aneuploidy occurs when there are errors in the specialized cell division of meiosis. Meiotic success is dependent on proper segregation of homologous chromosomes at meiosis I, and then subsequent sister chromatid segregation at meiosis II. There is still much that is unknown about the molecular mechanisms that contribute to infertility, and even less is known about the sexually dimorphic mechanisms during gametogenesis that can be contributing factors. The overall goal of the lab is to determine how these sexually dimorphic molecular mechanisms contribute to infertility. The goal of this proposal is to characterize the sex-specific roles of an enzyme crucial for relieving topological abnormalities in DNA during male and female meiosis. This enzyme, Topoisomerase II (Topo II), has been shown to have sex-specific phenotypes in both spermatogenesis and oogenesis in a variety of model organisms. Topo II also has roles in chromosome structure, condensation, and is a chromosome axis component in both mitosis and meiosis. Preliminary data indicates that Topo II also plays a role in the release of sister chromatid cohesion, an important step in ensuring the success of meiosis. The goals of this proposal will take a combined genetic, molecular biology, and biochemical approach to determine the sex-specific role of Topo II in meiosis. To achieve this, the applicant will be advised by a mentoring team that includes a sponsor with expertise in the fields of Meiosis and Genetics, as well as a co-sponsor who is an expert in Biochemistry, including protein expression, purification, and function. The collaboration between the candidate and her sponsors provides her with mentorship that is unique, interdisciplinary, and allows for the opportunity to develop a skillset that will be utilized throughout the entirety of her academic career, from post- doc to independent researcher. The resources, facilities, equipment, and faculty, available at the University of Delaware far exceed the needs of the proposal, ensuring a successful training environment to complete the aims proposed. Aim 1 uses single chromosome fluorescence labeling and nano-resolution microscopy to characterize the sexually dimorphic role of TOP-2 on sperm chromosome structure in late meiotic prophase l. I will also use a combination of genetic and confocal microscopy to determine the role of TOP-2 on oocyte chromosomes. Aim 2 will determine the differential protein interactions of TOP-2 in spermatogenesis and oogenesis using a biochemical approach in vivo and in vitro. These aims are innovative in the field of meiosis because the mechanisms known to govern the sex-specific differences in spermatogenesis and oogenesis are still largely unknown, exploring the role of Topo II will give us insight into one of the many functional and regulatory differences between male and female gametogenesis.

项目摘要 每个社会经济阶层的人，无论男女，每个民族和种族都受到影响 不孕不育非整倍体是导致不孕、出生缺陷和流产的主要原因。异倍 当减数分裂的特化细胞分裂出现错误时发生。减数分裂的成功取决于适当的 在减数分裂I时同源染色体分离，随后在减数分裂I时姐妹染色单体分离。 减数分裂II关于导致不孕的分子机制仍有许多未知之处， 甚至更少的是知道在配子发生过程中的性二型机制，可以促进 因素该实验室的总体目标是确定这些性二态分子机制 会导致不孕这项提议的目的是描述一种酶的性别特异性作用， 用于缓解雄性和雌性减数分裂期间DNA的拓扑异常。这种酶拓扑异构酶II (Topo II），已被证明在精子发生和卵子发生中具有性别特异性表型， 各种模式生物。Topo II还在染色体结构、凝聚中发挥作用，是一种 在有丝分裂和减数分裂中染色体轴的组成部分。初步数据显示，Topo II还发挥了 在释放姐妹染色单体凝聚中的作用，这是确保减数分裂成功的重要步骤。的 该提案的目标是采用遗传学、分子生物学和生物化学相结合的方法来确定 Topo II在减数分裂中的性别特异性作用。为了实现这一目标，申请人将由一个指导小组提供建议 其中包括一名在减数分裂和遗传学领域具有专业知识的赞助商，以及一名共同赞助商， 生物化学专家，包括蛋白质表达，纯化和功能。之间的协作 候选人和她的赞助商为她提供了独特的，跨学科的指导，并允许 有机会发展一套技能，将在整个学术生涯中使用，从后， 博士到独立研究员资源，设施，设备和教师，可在大学 特拉华州远远超出了建议的需要，确保成功的培训环境，以完成 提出的目标。AIM 1使用单染色体荧光标记和纳米分辨率显微镜来 TOP-2在减数分裂后期精子染色体结构中的性二态性作用。我 我们还将使用遗传学和共聚焦显微镜的组合来确定TOP-2对卵母细胞的作用 染色体目的二是确定TOP-2在精子发生中的差异蛋白相互作用， 在体内和体外使用生物化学方法进行卵子发生。这些目标在减数分裂领域是创新的 因为在精子发生和卵子发生中，控制性别特异性差异的机制是 仍然在很大程度上未知，探索Topo II的作用将使我们深入了解许多功能和 雄性和雌性配子发生的调节差异。