A MAP Kinase that controls the post-meiotic phase of the yeast life cycle.

一种控制酵母生命周期减数分裂后阶段的 MAP 激酶。

• 批准号: 10005813
• 负责人: JULIA Y LEE-SOETY
• 金额: $ 1.13万
• 依托单位: THOMAS JEFFERSON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-09-01 至 2020-09-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10005813
• 关键词: Address; Binding; Biochemical Genetics; Biological Models; C-terminal; Candidate Disease Gene; Catalytic Domain; Cell Nucleus; Cell division; Cells; ChIP-seq; Chromosome Segregation; Chromosomes; Clinical; Companions; Complex; Congenital Abnormality; DNA; DNA Polymerase II; DNA biosynthesis; DNA-Binding Proteins; DNA-Directed RNA Polymerase; Data; Data Set; Defect; Development; Diploid Cells; Diploidy; Engineering; Epitopes; Equation; Event; Family member; Follow-Up Studies; Gametogenesis; Genes; Genetic; Genetic Recombination; Genetic Transcription; Germ Cells; Haploid Cells; Haploidy; Health; Home environment; Homologous Gene; Human; Immunofluorescence Immunologic; Infertility; Institution; Laboratories; Life Cycle Stages; MAP Kinase Gene; Malignant Neoplasms; Meiosis; Methods; Mitogen-Activated Protein Kinases; Mitosis; Modification; Molecular Genetics; Monitor; Morphogenesis; Mothers; Mutation; Names; Nuclear; Nuclear Accidents; Nutrient; Organism; Pathway interactions; Pattern; Permeability; Phase; Philadelphia; Phospho-Specific Antibodies; Phosphorylation; Phosphotransferases; Principal Investigator; Process; Promoter Regions; Prophase; Proteins; RNA; RNA Polymerase II; Reproduction spores; Research; S Phase; Saccharomyces cerevisiae; Saints; Serine; Spontaneous abortion; Starvation; Students; System; Testing; Training; Universities; Work; Yeast Model System; Yeasts; analog; bioinformatics tool; crosslink; developmental disease; genetic approach; inhibitor/antagonist; insight; macromolecule; programs; promoter; purine analog; recruit; response; sabbatical; segregation; spatiotemporal; undergraduate research

Project Summary/Abstract: Meiosis is the specialized form of cell division that generates haploid gametes from diploid precursors. It is essential for the life-cycles of almost all eukaryotic organisms. Understanding meiotic regulatory mechanisms is relevant to human health since defects in meiosis cause infertility, cancer, and birth defects. Baker's yeast is an outstanding model system to study meiotic development. In response to starvation, diploid cells enter meiotic development and generate four haploid cells within the mother cell. Early events include meiotic DNA replication, homolog pairing, and genetic recombination. After the completion of genetic recombination, cells undergo 2 rounds of chromosome segregation without an intervening S-phase. After the second meiotic division (MII), the haploid products are encased in spore walls that protect the haploids from environmental insults. Although yeast has taught us much about meiotic S-phase, prophase, and the meiotic divisions, much less is known about the processes that occur after MII has taken place. One reason we know so little about the post-meiotic phase of this program is that the spore walls are experimentally impenetrable; extracting macromolecules under native conditions or monitoring molecules using immunofluorescence methods has until recently been nearly impossible. Previous work in the sponsor's laboratory has identified a meiosis-specific MAPK named Smk1 that controls the post-meiotic program of spore formation. This led to the discovery that Smk1 activates the expression of late meiosis-specific genes, which are uniquely expressed after the spore wall has assembled. This proposal takes advantage of engineered yeast strains that produce permeable spore walls so that the intracellular and nuclear events that occur during the late stages of meiotic development can be studied. The proposal also takes advantage of engineered forms of Smk1 that are sensitive to cell-permeable purine analogs (smk1-as) that can be used to turn Smk1 on and off. Mutations that permeabilize the spore wall with smk1-as are combined in a genetic background that undergoes meiosis in a highly synchronous manner, and this will be used to define the complete set of late meiosis-specific genes that are regulated by Smk1 (aim 1). We will also test the hypothesis that Smk1 is recruited to late meiosis-specific transcriptional promoters (aim 2). Preliminary data suggests that Smk1 phosphorylates the C terminal domain of RNA polymerase, and we will also test this hypothesis and whether these modifications take place at late meiosis specific genes as they are being transcribed (aim 3). Studies of how meiotic events are regulated in yeast may provide insights into human gametogenesis, which have clinical implications in solving infertility, miscarriages, and developmental disorders. The principal investigator (Julia Lee-Soety) intends to conduct this work during her sabbatical leave in Dr. Edward Winter's laboratory at the Thomas Jefferson University in Philadelphia. Beyond the project period, the PI plans to continue follow-up studies with undergraduate research students at her home institution (Saint Joseph's University).

项目概要/摘要：减数分裂是产生单倍体配子的细胞分裂的特殊形式 从二倍体前体。它对几乎所有真核生物的生命周期都是必不可少的。理解 减数分裂调节机制与人类健康有关，因为减数分裂缺陷会导致不育，癌症， 出生缺陷。面包酵母是研究减数分裂发育的一个优秀的模式系统。响应于 在饥饿时，二倍体细胞进入减数分裂发育并在母细胞内产生四个单倍体细胞。早期 事件包括减数分裂DNA复制、同源配对和遗传重组。完成后 在遗传重组中，细胞经历2轮染色体分离而没有居间的S期。 在第二次减数分裂（MII）后，单倍体产物被包裹在孢子壁中， 从环境的侮辱。虽然酵母已经教会了我们很多关于减数分裂S期，前期， 减数分裂，更少的是知道的过程后发生的MII已经发生。一 我们对这个程序的减数分裂后阶段知之甚少的原因是孢子壁是实验性的。 不可穿透;在天然条件下提取大分子或使用 免疫荧光方法直到最近几乎是不可能的。以前在赞助商的 一个实验室已经鉴定出一种名为Smk 1的减数分裂特异性MAPK，它控制孢子的减数分裂后程序 阵这导致了Smk 1激活晚期减数分裂特异性基因的表达的发现， 是在孢子壁组装后唯一表达的。该提案利用了工程设计的优势 酵母菌株产生可渗透的孢子壁，从而使细胞内和细胞核事件发生在 可以研究减数分裂发育的后期阶段。该提案还利用了工程 Smk 1的形式，对可用于开启Smk 1的细胞渗透性嘌呤类似物（smk 1-as）敏感 然后离开用smk1-as渗透孢子壁的突变在遗传背景中结合， 以高度同步的方式进行减数分裂，这将被用来定义一套完整的晚期减数分裂。 减数分裂特异性基因，由Smk1（目的1）。我们还将检验Smk 1是 募集到后期减数分裂特异性转录启动子（AIM 2）。初步数据表明，Smk 1 磷酸化RNA聚合酶的C末端结构域，我们也将测试这一假设， 这些修饰发生在减数分裂后期特定基因转录时（aim 3）。研究 酵母中减数分裂事件的调节方式可能为人类配子发生提供见解， 在解决不孕症、流产和发育障碍方面的意义。首席研究员（Julia） Lee-Soety）打算在她的休假期间在爱德华·温特博士的实验室进行这项工作。 费城的托马斯杰斐逊大学。在项目期后，PI计划继续随访 在她的家乡机构（圣约瑟夫大学）与本科研究生一起学习。