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

项目摘要 减数分裂是一种专门的细胞分裂，其特征是一轮DNA复制，然后是两轮 染色体隔离，导致游戏的形成。粘着蛋白是染色体相关的 维持姐妹染色质凝聚力的多蛋白环，这对于准确 染色体/染色单体分离。在减数分裂过程中，粘着蛋白拆卸特别复杂 在第一个减数分裂师（MI）和 然后在第二个减数分裂师（MII）期间的中心码。 NIMA样激酶1（NEK1）是双重特异性 在生殖细胞中高度表达的丝氨酸/苏氨酸和酪氨酸激酶。小鼠中NEK1的损失导致 保留在MI处的染色体臂上的粘蛋白亚基SMC3及随后的不育症。粘着蛋白去除 首先由“预言途径”策划两个步骤，然后是分离酶介导的裂解 粘着蛋白环。预言途径是由翅膀 - 易于式蛋白质之间的化学计量定义的 （WAPL）和索罗林（Sororin 它在减数分裂中描述。我的初步数据揭示了NEK1在粘蛋白调节中的令人兴奋的作用 MI处的动力学，直接在粘蛋白亚基，SMC3，RAD21L和REC8的水平上，并间接地 通过磷酸化PDS5B-WAPL复合物。此外，我的研究表明NEK1动作 在预言途径上是通过蛋白质磷酸酶1-伽马（PP1γ）介导的，这是磷酸的 NEK1，并结合并去磷酸化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中执行其他角色，这些角色不同于其在其上的活性 凝聚力动力学。那我假设NEK1充当减数分裂事件的主要调节剂，主要是 通过其对关键组成部分的作用 预言途径，但也在策划其他NEK激酶的作用时。我的长期目标是阐明 NEK1如何调节MI的凝聚力去除，但也进一步表征了NEK1催化活性的作用 在两个减数分裂师的策划下游事件时。提出了三个具体目标：（1）到 阐明NEK1在PP1γ磷酸化中的作用，（2）评估WAPL的重要性 在MI和（3）的内聚力去除时磷酸化，以阐明NEK1作为主调节剂的功能 减数分裂期间的Nek家族。这些实验将提供一个新颖而令人兴奋的数据，以描述 第一次，NEK1在减数分裂预言途径期间在调节粘蛋白中的调节中的作用。 申请人Brieño-Enríquez博士是医学博士学位，在配子生物学领域具有考虑经验。 申请人研究了人类和小鼠中配子生物学的独特过程，目的是了解 为有性繁殖带来健康游戏的复杂事件。最终目标 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 哺乳动物的减数分裂和配子发生受调节，哪些事件在人类中可能有缺陷， 与减数分裂相关的错误率异常高。那是一个职业发展计划 旨在为Brieño-Enríquez博士提供建立强大研究计划的所有必要工具 除了可用的最佳科学资源外，Brieño-Enríquez博士还将 从他的主办机构康奈尔大学（Cornell University）提供的各种讲习班和半小伙子中受益。 主题包括求职，准备申请和准备面试，谈判初创公司 包装，建立实验室并建立一个有效的研究团队。此外，还有专门的课程 提高教学技巧，写作补助以及处理工作场所的冲突，仅提及一些。博士 Brieño-Enríquez还将受益于由他的精神的杰出心理团队Paula Cohen博士 马克·罗伯森（Mark Roberson） 具有心理成功的博士后研究员的经验。支持这支心理团队是一个杰出的 咨询委员会由激酶生物学领域的专家（Marcus Smolka博士），粘粘蛋白（博士）组成（博士 迈克尔·戈德堡（Michael Goldberg），小鼠转基因和繁殖（John Schimenti博士）。申请人将开会 经常与他的导师和咨询团队非正式和正式。而且，导师和 咨询委员会致力于协助布里诺·埃里克斯（Brieño-Enríquez）博士发展研究技能，并 将帮助他制作强大的教师申请，工作精神和粉笔谈话。最后，科恩博士承诺 通过他的研究在财务上为Brieño-Enríquez博士提供支持，并在他的情况下提供持续的心态 过渡到他新独立的立场。 总而言之，Brieño-Enríquez博士设计了一个令人兴奋而新颖的建议，旨在定义角色 减数分裂细胞分裂中的NIMA样激酶。这样，他将为 他自己在配子生物学领域。为了协助他，他组建了一个世界一流的心理团队和一个 强大的发展计划，所有这些目标都是统一的目标，即促进成功过渡到独立性 令人兴奋的年轻科学家