The role of PAM-1 in the regulation of the cell cycle and polarity establishment in C. elegans

PAM-1 在秀丽隐杆线虫细胞周期和极性建立调节中的作用

基本信息

批准号：
10359983
负责人：
REBECCA LYNN LYCZAK
金额：
$ 40.79万
依托单位：
URSINUS COLLEGE
依托单位国家：
美国
项目类别：
财政年份：
2014
资助国家：
美国
起止时间：
2014-09-24 至 2024-08-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10359983
关键词：
Affect Anterior CDC2 gene Caenorhabditis elegans Cell Cycle Cell Cycle Regulation Cell Cycle Stage Cell Polarity Cells Complex Coupled Cyclin B Cyclin-Dependent Kinases Cyclins Development Developmental Process Embryo Fertility Genes Goals Homologous Gene Image Infertility Lesion Maintenance Maturation-Promoting Factor Meiosis Mentors Metalloproteases Mitosis Modeling Molecular Mutate Mutation Oocytes Organism Phenotype Phosphoric Monoester Hydrolases Phosphotransferases Play Process Proteins Publishing RNA Interference Regulation Role STEM field Science Series Students System Testing Time Tissues Underrepresented Students Work cancer prevention career causal variant embryo cell experimental study genome sequencing imaging genetics meetings mutant novel oocyte maturation puromycin-sensitive aminopeptidase student mentoring success tissue regeneration undergraduate student whole genome

项目摘要

PROJECT SUMMARY The cell cycle is a tightly regulated process controlled by the transient activation of cyclin- dependent kinases (CDKs). Activation and deactivation of CDKs is coordinated by its associated cyclin as well as other kinases, phosphatases, and the degradation machinery. Coordination of these complexes drives the cell through each stage of the cell cycle and is necessary for development, tissue maintenance, and fertility. Puromycin-sensitive aminopeptidases (PSAs) are highly conserved metalloproteases implicated in cell-cycle regulation in numerous organisms. Despite widespread roles in this process, the mechanism by which PSAs interact with the cyclin/CDKs and other cell-cycle machinery is poorly understood. A key goal of our project is to uncover these interactions in the C. elegans model. In our previous work, we showed that the C. elegans PSA homolog, PAM-1, plays a role in meiotic exit regulation and anterior-posterior axis establishment and when mutated, results in embryonic lethality. In a screen for suppressors of pam-1, we identified a mutation in wee-1.3 that rescues the lethality of pam-1 mutants. WEE-1.3 is an inhibitory kinase that negatively regulates CDK-1, part of the maturation promoting complex. We went on to show that pam-1 and wee-1.3 genetically interact in polarity establishment and oocyte maturation, suggesting a broad role for PAM-1 in regulating the cell cycle. The proposed work seeks to further characterize the role of PAM-1 and its interaction with WEE-1.3 and the MPF, testing the hypothesis that PAM-1 is necessary for full MPF activity. Work will focus on characterization of oocyte maturation, meiosis, and mitosis in the early embryo through a combination of time-lapse imaging, genetic interaction, and localization studies. In addition, we will characterize and identify the genes mutated in additional suppressors of pam-1 to find new interactions. We expect to gain a new understanding of how PAM-1 regulates that cell cycle that is likely to be applicable to other systems. The proposed experiments will be largely carried out by undergraduates who will be mentored to provide them the guidance and expertise needed for success in science careers and graduate work. Inclusion and mentoring of students underrepresented in STEM is an important component of this work.

项目概要细胞周期是一个严格调控的过程，由细胞周期蛋白的瞬时激活控制。依赖性激酶（CDK）。 CDK 的激活和停用由其协调相关的细胞周期蛋白以及其他激酶、磷酸酶和降解机制。这些复合物的协调驱动细胞经历细胞周期的每个阶段，发育、组织维持和生育所必需的。嘌呤霉素敏感氨肽酶 (PSA) 是与细胞周期有关的高度保守的金属蛋白酶许多生物体中的调节。尽管在此过程中发挥了广泛的作用，但该机制哪些 PSA 与细胞周期蛋白/CDK 和其他细胞周期机制相互作用尚不清楚。我们项目的一个关键目标是揭示线虫模型中的这些相互作用。在我们的在之前的工作中，我们表明秀丽隐杆线虫 PSA 同源物 PAM-1 在减数分裂退出中发挥作用调节和前后轴的建立，当突变时，会导致胚胎杀伤力。在筛选 pam-1 抑制剂时，我们发现 wee-1.3 中的一个突变可以挽救 pam-1突变体的致死率。 WEE-1.3 是一种抑制性激酶，负向调节 CDK-1，成熟促进复合体的一部分。我们继续证明 pam-1 和 wee-1.3 在极性建立和卵母细胞成熟中遗传相互作用，表明 PAM-1 调节细胞周期。拟议的工作旨在进一步描述 PAM-1 及其与 WEE-1.3 和 MPF 的相互作用，检验 PAM-1 的假设全面强积金活动所必需的。工作重点是卵母细胞成熟的表征，通过结合延时成像、遗传技术来观察早期胚胎的减数分裂和有丝分裂交互和本地化研究。此外，我们还将表征和鉴定基因在 pam-1 的其他抑制子中发生突变以发现新的相互作用。我们期望获得新的了解 PAM-1 如何调节细胞周期，这可能适用于其他细胞周期系统。拟议的实验将主要由本科生进行，他们将接受指导，为他们提供科学事业成功所需的指导和专业知识和毕业作品。对 STEM 中代表性不足的学生进行包容和指导是一项这项工作的重要组成部分。