Evolutionary adaptation and spatial organization of signaling in the Mitotic Exit Network

项目摘要 通过信号级联来感知和处理信息是细胞生命的重要组成部分。几个 在真核生物中，MAP激酶和河马通路等信号级联信号仍然普遍存在。 在不同的生物体中执行不同的功能。尽管人们对这些途径进行了充分的研究，但它们是如何进化到 承担新的职能和适应新的投入仍然知之甚少。有丝分裂退出网络(MEN)，a Ras样GTPase信号级联和河马途径的酵母同源物，提供了一个独特的机会 研究这个问题。在男性中，相同的核心信号组件以不同的方式在 不同的发展轨迹。在酵母有丝分裂期间，通过不对称的细胞分裂发生 这些人被称为萌芽，被固定在纺锤体的极体上(SPB，相当于酵母的中心体)。 并通过其GTP酶TEM1对纺锤体位置做出反应。在减数分裂期间，萌芽被抑制， 因此，不需要感知心轴位置，SPBS不再组织男性信号，而且不清楚 男性激活是否仍然需要TEM1，以及它可能对什么信号做出反应。要了解 男性在不同细胞环境下的适应，这一提议将检验这种适应的假设 部分由MEN激酶CDC15，Tem1的效应蛋白激酶， 在有丝分裂和减数分裂之间。新冠肺炎疫情相关研究限制与导师逝世 在大流行期间，Angelika Amon博士严重扰乱了我的研究进展，延误了我的职业生涯 发展计划。虽然我在实现我最初的K99提案中提出的目标方面取得了重大进展， 我目前正在准备一个关于这个项目的手稿，我向独立的过渡推迟了一年 相对于最初提议的时间表。资助的延期将使我能够发展蛋白质方面的关键技能 生物化学、体外重建和酵母减数分裂以剖析CDC15调控的机制。 有丝分裂与减数分裂。我将在我的导师斯蒂芬·贝尔博士的指导和培训下完成这一目标 (生物化学，单分子，和体外重建)和ElçinÜnual博士(酵母减数分裂，我的成员 咨询委员会)。此外，提供资金的扩展还将使我能够在 确保获得学术职位，并使我能够过渡到一名独立调查员。总而言之， 我将在延长的K99期间接受额外的培训和支持，这将使我具备以下知识 以及研究不同细胞中男性信号适应的潜在机制所需的技能 并为理解进化论的独立研究生涯奠定了坚实的基础 细胞信号的适应性。

Project Summary Sensing and processing information through signaling cascades is an essential part of cellular life. A few signaling cascades such as the MAP kinase and Hippo pathways are ubiquitous among eukaryotes yet perform different functions across organisms. Although these pathways are well-studied, how they evolve to take on new functions and adapt to new inputs remains poorly understood. The Mitotic Exit Network (MEN), a Ras-like GTPase signaling cascade and yeast homolog of the Hippo pathway, provides a unique opportunity to study this question. In the MEN, the same core signaling components operate in distinct manners under different developmental trajectories. During yeast mitosis which occurs through an asymmetric cell division called budding, the MEN is scaffolded onto the spindle pole bodies (SPB, the yeast equivalent of centrosomes) and responds to spindle position through its GTPase Tem1. During meiosis, where budding is suppressed and thus no need to sense spindle position, MEN signaling is no longer organized at the SPBs, and it is unclear whether Tem1 is still required for MEN activation and what signal it may respond to. To understand the adaptation of the MEN under distinct cellular contexts, this proposal will test the hypothesis that this adaptation is enabled partially by different activation mechanisms of the MEN kinase Cdc15, the effector kinase of Tem1, between mitosis and meiosis. COVID-19 pandemic related research restriction and the passing of my mentor Dr. Angelika Amon during the pandemic severely disrupted my research progress and delayed my career development plans. While I made significant progress toward the aims laid out in my original K99 proposal and am currently preparing a manuscript on the project, my transition to independence was delayed for a year relative to the originally proposed timeline. A funded extension will allow me to develop critical skills in protein biochemistry, in vitro reconstruction, and yeast meiosis to dissect the mechanisms of Cdc15 regulation in mitosis versus meiosis. I will accomplish this with the guidance and training from my mentor Dr. Stephen Bell (biochemistry, single molecule, and in vitro reconstruction) and Dr. Elçin Ünal (yeast meiosis, member of my advisory committee). Furthermore, a funded extension will also allow me to complete my current search in securing an academic position and enable my transition into an independence investigator. In summary, the additional training and support I will receive during the extended K99 period will equip me with the knowledge and skills necessary to study the mechanisms underlying signaling adaptation of the MEN in different cellular contexts and create a strong foundation for an independent research career in understanding the evolutionary adaptation of cellular signaling.