NIMA-like Kinase NEK1 as a Regulator of Mammalian Gametogenesis

NIMA 样激酶 NEK1 作为哺乳动物配子发生的调节剂

• 批准号: 9902491
• 负责人: MIGUEL ANGEL BRIENO-ENRIQUEZ
• 金额: $ 17.49万
• 依托单位: MAGEE-WOMEN'S RES INST AND FOUNDATION
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-03-27 至 2022-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9902491
• 关键词: Advisory Committees; Affect; Alanine; Aneuploidy; Area; Aspartic Acid; Binding; Binding Proteins; Biochemical; Biological Assay; Biology; CRISPR/Cas technology; CSPG6 gene; Catalytic Domain; Cell division; Centromere; Centrosome; Chalk; Chromatids; Chromosome Arm; Chromosome Pairing; Chromosome Segregation; Chromosomes; Complex; Conflict (Psychology); Congenital Abnormality; Coupled; Cytology; DNA biosynthesis; Data; Development Plans; Doctor of Philosophy; Educational process of instructing; Educational workshop; Event; Excision; Faculty; Family; Family member; G2/M Transition; Gametogenesis; Gene Transfer Techniques; Genes; Genetic Crossing Over; Germ Cells; Glutamic Acid; Goals; Grant; Histology; Human; Immunoprecipitation; In Vitro; Infertility; Institution; Interview; Laboratories; Lead; Mediating; Mediation; Meiosis; Meiotic Prophase I; Mentors; Mentorship; Methods; Microtubules; Mitosis; Mus; Mutant Strains Mice; NIMA; Occupations; Pathway interactions; Phase; Phenotype; Phosphoric Monoester Hydrolases; Phosphorylation; Phosphotransferases; Play; Population; Positioning Attribute; Postdoctoral Fellow; Premature Ovarian Failure; Process; Prophase; Protein Tyrosine Kinase; Protein phosphatase; Protein-Serine-Threonine Kinases; Proteins; Proteomics; Regulation; Reproduction; Reproductive Biology; Research; Resources; Role; Scientist; Serine; Signal Pathway; Sister Chromatid; Somatic Cell; Spontaneous abortion; Testing; Time; United States; Universities; Variant; Wing; Workplace; Writing; career development; cohesin; cohesion; experience; experimental study; human error; improved; member; mutant; novel; premature; programs; protein complex; reproductive; segregation; separase; skills; stoichiometry; tool

Project summary Meiosis is a specialized cell division characterized by a single round of DNA replication followed by two rounds of chromosome segregation, resulting in the formation of gametes. Cohesin is a chromosome-associated multiprotein ring that maintains sister chromatid cohesion, and which is essential for accurate chromosome/chromatid segregation. During meiosis, cohesin disassembly is particularly complicated by the requirement for sequential loss of cohesion along the chromosome arms at the first meiotic division (MI) and then at the centromere during the second meiotic division (MII). NIMA-like kinase 1 (NEK1) is a dual specific serine/threonine and tyrosine kinase that is highly expressed in germ cells. Loss of NEK1 in mice leads to retention of the cohesin subunit SMC3 on chromosome arms at MI and subsequent infertility. Cohesin removal is orchestrated in two steps, first by “the prophase pathway”, followed by Separase-mediated cleavage of the cohesin ring. The prophase pathway is defined by the stoichiometry between the Wings-apart-like protein (WAPL) and Sororin, which compete for binding to PDS5B on the cohesin ring, but this pathway has not yet been described in meiosis. My preliminary data reveal exciting roles for NEK1 in the regulation of cohesin dynamics at MI, both directly at the level of the cohesin subunits, SMC3, RAD21L and REC8, and indirectly through phosphorylation the PDS5B-WAPL complex. Furthermore, my studies have shown that NEK1 action on the prophase pathway is mediated via Protein Phosphatase 1-gamma (PP1γ), which is a phosphotarget of NEK1, and which binds and de-phoshphorylates WAPL. Moreover, my preliminary studies indicate that NEK1 also regulates a cascade of other NEK proteins to perform other roles in MI that are distinct from its activity on cohesion dynamics. Thus, I hypothesize that NEK1 acts as master regulator of events in meiosis, primarily playing a crucial role in the timing of cohesin removal at MI through its actions on critical components of the prophase pathway, but also in orchestrating the actions of other NEK kinases. My long term goal is to elucidate how NEK1 regulates cohesion removal at MI, but also to further characterize the role of NEK1 catalytic activity in orchestrating downstream events at both meiotic divisions. Three specific aims are proposed: (1) To elucidate the role of NEK1 in the phosphorylation of PP1γ, (2) To assess the importance of WAPL phosphorylation on cohesion removal at MI and (3) To elucidate the function of NEK1 as a master regulator of the NEK family during meiosis. These experiments will provide a novel and exciting data that describes, for the first time, the the role of NEK1 in the regulation of cohesin removal during the prophase pathway in meiosis. The applicant, Dr. Brieño-Enríquez, is an MD, PhD with considerable experience in the field of gamete biology. The applicant has studied distinct process of gamete biology in humans and mice, with a goal to understanding the complex events that give rise to healthy gametes for sexual reproduction. The ultimate goal of the applicant is to establish a unique and state-of-the-art research program aimed at understanding how mammalian meiosis and gametogenesis is regulated and what events may be defective in humans, where the error rates associated with meiosis are exceptionally high. Thus, a career development plan has been established to provide Dr. Brieño-Enríquez with all the necessary tools to establish a robust research program in a stellar academic institution. In addition to the best scientific resources available, Dr. Brieño-Enríquez will benefit from a wide variety of workshops and seminars available at his host institution, Cornell University. Topics include job searches, preparing an application and preparing for an interview, negotiating a startup package, establishing a lab, and building an effective research team. In addition, there are courses devoted to improving teaching skills, writing grants, and dealing with conflict in the work place, to mention just a few. Dr. Brieño-Enríquez will also benefit from an outstanding mentoring team, led by his mentors, Dr. Paula Cohen and Dr. Mark Roberson, both highly regarded in the field of Reproductive Biology, and both having much experience in mentoring successful postdoctoral fellows. Supporting this mentoring team is an outstanding advisory committee consisting of experts in the field of kinase biology (Dr. Marcus Smolka), cohesins (Dr. Michael Goldberg), mouse transgenesis and reproduction (Dr. John Schimenti). The applicant will meet frequently with his mentors and advisory team, both informally and formally. Moreover, the mentors and advisory committee have committed to assisting Dr. Brieño-Enríquez as he develops his research skills, and will help him to craft a strong faculty application, job seminar, and chalk talk. Lastly, Dr. Cohen has committed to supporting Dr. Brieño-Enríquez financially through his research and to providing ongoing mentorship as he transitions to his newly independent position. In summary, Dr. Brieño-Enríquez has devised an exciting and novel proposal that seeks to define the role of the NIMA-like kinases in the meiotic cell divisions. In doing so, he will create a robust and novel niche for himself in the field of gamete biology. To assist him, he has assembled a world-class mentoring team and a strong development plan, all with a unified goal of promoting the successful transition to independence of this exciting young scientist

项目概要 减数分裂是一种特殊的细胞分裂，其特征是单轮 DNA 复制，然后是两轮 染色体分离，导致配子的形成，粘连蛋白是染色体相关的。 维持姐妹染色单体凝聚力的多蛋白环，这对于准确 染色体/染色单体分离在减数分裂过程中，粘连蛋白的分解尤其复杂。 第一次减数分裂 (MI) 时沿染色体臂连续丧失内聚力的要求和 然后在第二次减数分裂 (MII) 期间的着丝粒处，NIMA 样激酶 1 (NEK1) 是一种双特异性激酶。 小鼠生殖细胞中高表达的丝氨酸/苏氨酸和酪氨酸激酶 NEK1 缺失会导致小鼠死亡。 MI 时，粘连蛋白亚基 SMC3 保留在染色体臂上，随后去除粘连蛋白。 分两步精心策划，首先是“前期途径”，然后是分离酶介导的裂解 粘连蛋白环。前期途径由翼分离样蛋白之间的化学计量定义。 (WAPL) 和 Sororin，竞争结合粘连蛋白环上的 PDS5B，但该途径尚未 我的初步数据揭示了 NEK1 在粘连蛋白调节中的令人兴奋的作用。 MI 的动力学，直接在粘连蛋白亚基、SMC3、RAD21L 和 REC8 水平，以及间接 通过磷酸化 PDS5B-WAPL 复合物 此外，我的研究表明 NEK1 具有作用。 前期途径是通过蛋白磷酸酶 1-gamma (PP1γ) 介导的，它是 NEK1，它结合并去磷酸化 WAPL 此外，我的初步研究表明 NEK1。 还调节其他 NEK 蛋白的级联，以在 MI 中发挥与其活性不同的其他作用 因此，我认为 NEK1 主要是减数分裂事件的主要调节因子。 通过对 MI 的关键组成部分的作用，在 MI 的粘连蛋白去除时间中发挥着至关重要的作用 我的长期目标是阐明前期途径，而且还参与协调其他 NEK 激酶的作用。 NEK1 如何调节 MI 中的内聚力去除，同时进一步表征 NEK1 催化活性的作用 提出了三个减数分裂下游事件的具体目标：（1） 阐明 NEK1 在 PP1γ 磷酸化中的作用，(2) 评估 WAPL 的重要性 (3) 阐明 NEK1 作为 MI 处内聚力去除的主要调节因子的功能 这些实验将提供描述减数分裂期间 NEK 家族的新颖且令人兴奋的数据。 首次揭示了 NEK1 在减数分裂前期途径中粘连蛋白去除调节中的作用。 申请人Brieño-Enríquez博士是一位医学博士、博士，在配子生物学领域拥有丰富的经验。 申请人研究了人类和小鼠配子生物学的不同过程，目的是了解 产生健康配子以进行有性生殖的复杂事件。 申请人将建立一个独特且最先进的研究计划，旨在了解如何 哺乳动物减数分裂和配子发生受到调节，哪些事件在人类中可能存在缺陷，其中 与减数分裂相关的错误率非常高，因此，职业发展计划已经制定。 成立的目的是为 Brieño-Enríquez 博士提供建立强有力的研究计划所需的所有工具 除了拥有最好的科学资源外，Brieño-Enríquez 博士还将在一流的学术机构中学习。 受益于他的主办机构康奈尔大学举办的各种研讨会和研讨会。 主题包括求职、准备申请和准备面试、谈判初创公司 此外，还有专门的课程。 提高教学技能、撰写补助金以及处理工作场所的冲突，仅举几例。 Brieño-Enríquez 还将受益于由他的导师 Paula Cohen 博士领导的优秀指导团队 和马克·罗伯森博士，两人在生殖生物学领域都享有很高的声誉，并且都有很多 指导成功的博士后研究员的经验支持这个指导团队是一个杰出的。 顾问委员会由激酶生物学领域的专家（Marcus Smolka 博士）、粘连蛋白（Dr. Marcus Smolka）组成。 Michael Goldberg），小鼠转基因和繁殖（John Schimenti 博士）。 经常与他的导师和顾问团队进行非正式和正式的交流。 顾问委员会致力于协助 Brieño-Enríquez 博士发展研究技能，并且 将帮助他撰写一份强有力的教师申请、工作研讨会和粉笔演讲。最后，科恩博士做出了承诺。 通过 Brieño-Enríquez 博士的研究提供经济支持，并在他的研究过程中提供持续的指导 过渡到他新独立的职位。 总之，Brieño-Enríquez 博士提出了一项令人兴奋且新颖的提案，旨在定义 减数分裂细胞分裂中的 NIMA 样激酶在此过程中，他将创造一个强大而新颖的利基市场。 为了帮助他自己在配子生物学领域的发展，他组建了一个世界级的指导团队和一个团队。 强有力的发展计划，所有这些都有一个统一的目标，即促进该国成功过渡到独立 令人兴奋的年轻科学家