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.

项目总结 每个社会经济阶层的人，包括男性和女性，以及每个民族和族裔，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。非整倍体 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 同源染色体在减数分裂I时分离，然后姐妹染色单体在减数分裂I时分离 减数分裂II.关于导致不孕症的分子机制仍有许多未知之处，以及 对配子发生过程中可能起作用的性二态机制知之甚少 各种因素。该实验室的总体目标是确定这些性二态分子机制是如何 导致不孕不育。这项提议的目标是描述一种关键酶的性别特异性作用。 用于缓解雄性和雌性减数分裂过程中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都具有性别特异性表型。 各种模式生物。Topo II也在染色体结构、凝聚中发挥作用，是一种 有丝分裂和减数分裂中的染色体轴成分。初步数据显示，Topo II也起到了 在姐妹染色单体凝聚力的释放中发挥作用，这是确保减数分裂成功的重要一步。这个 这项提案的目标将采用遗传学、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。为了做到这一点，申请人将得到一个指导团队的建议 这包括一位在减数分裂和遗传学领域具有专业知识的赞助商，以及一位是 生物化学方面的专家，包括蛋白质的表达、纯化和功能。两家公司之间的合作 候选人和她的赞助人为她提供了独特的、跨学科的指导，并允许 有机会培养一种技能，这种技能将在她的整个学术生涯中使用，从后 医生给独立研究人员。牛津大学提供的资源、设施、设备和师资 特拉华州远远超出了提案的需求，确保了成功的培训环境，以完成 目标建议。目标1使用单染色体荧光标记和纳米分辨率显微镜来 L减数分裂晚期前期精子染色体结构中TOP-2的性别二态作用 还将结合使用遗传显微镜和共聚焦显微镜来确定TOP-2在卵母细胞上的作用 染色体。目的2将确定TOP-2在精子发生和发育过程中的差异蛋白质相互作用 利用体内和体外的生物化学方法进行卵子发生。这些目标在减数分裂领域具有创新性。 因为已知的控制精子发生和卵子发生性别差异的机制是 在很大程度上仍不为人所知，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控差异。