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

项目摘要 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（CDKs）。CDK的激活和失活是由其 相关的细胞周期蛋白以及其他激酶、磷酸酶和降解机制。 这些复合物的配位驱动细胞通过细胞周期的每个阶段， 是发育、组织维持和生育所必需的。嘌呤霉素敏感 氨肽酶（PSAs）是高度保守的参与细胞周期的金属蛋白酶 在许多生物体中的调节。尽管在这一过程中发挥着广泛的作用， 对哪些PSA与细胞周期蛋白/CDK和其它细胞周期机制相互作用知之甚少。 我们项目的一个关键目标是揭示C中的这些相互作用。elegans模型在我们 以前的工作，我们表明，C。秀丽线虫PSA同源物PAM-1在减数分裂退出中起作用 调节和前后轴的建立，当突变时，导致胚胎 杀伤力在pam-1抑制基因的筛选中，我们在wee-1.3中发现了一个突变， PAM-1突变体的致死性。WEE-1.3是负调节CDK-1的抑制性激酶， 成熟促进复合物的一部分。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的遗传相互作用，这表明 PAM-1在细胞周期调控中的作用。拟议的工作旨在进一步确定 3和MPF的相互作用，检验PAM-1是一种新的蛋白质的假设。 全面开展强积金活动。工作将集中在卵母细胞成熟的表征， 减数分裂和有丝分裂在早期胚胎通过组合的时间推移成像，遗传 互动和本地化研究。此外，我们将描述和识别基因 在PAM-1的另外的抑制子中突变以发现新的相互作用。我们希望获得一个新的 了解PAM-1如何调节细胞周期，这可能适用于其他 系统.拟议中的实验将主要由本科生进行，他们将 指导他们为科学事业的成功提供指导和专业知识 毕业工作。包容和指导在STEM中代表性不足的学生是一个 这项工作的重要组成部分。

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.