Genetic and Molecular Dissection of Regulatory Mechanisms Underlying Temperature and Nutritional Compensation of the Circadian Clock in Neurospora crassa

项目摘要/摘要： 生物钟提供计时功能，以预测环境的日常变化，并被使用 地球上几乎所有的真核生物。~24小时的昼夜节律周期长度针对波动进行了缓冲 在外部条件下，包括温度和营养水平，在一种普遍存在的现象中称为 补偿。补偿的分子机制在任何真核生物中都不是很清楚。 有机体。该项目的长期目标是了解温度补偿(TC)、营养 补偿(NC)，以及补偿效应器和核心时钟之间的分子相互作用 网络使用真核生物粗糙脉孢菌的模型。尽管研究和培训显著地 被新冠肺炎大流行扰乱，TC和NC缺陷突变的反向基因筛查 使用脉孢子虫基因敲除集合完成(目标1)。具有NC缺陷的突变体的数量为 由于这项工作，增加了一倍以上。鉴定TC相关物质的质谱学实验 酪蛋白激酶1和酪蛋白激酶2的底物研究进展顺利，预计将于2022年发表(目标2)。 这一行政副刊将支持因 新冠肺炎大流行以及这一奖学金下的第三个第一作者手稿的出版。 由于研究的完成，昼夜节律补偿的分子机制开始形成 在这个项目下。这个和未来的独立调查项目将解决~24小时昼夜节律 研究发现，在环境中营养水平和温度不同的情况下，周期长度是保持不变的 这对患有昼夜节律睡眠障碍的人类、倒班工人、持续的时差反应以及其他 时间生物学条件。

Project Summary/Abstract: The circadian clock serves a time-keeping function to anticipate daily changes in the environment and is used by almost every eukaryote on the planet. The ~24-hour circadian period length is buffered against fluctuations in external conditions, including temperature and nutrient levels, in a ubiquitous phenomenon called compensation. The molecular mechanism underlying compensation is not well understood in any eukaryotic organism. The long-term goal of this project is to understand temperature compensation (TC), nutritional compensation (NC), and the molecular interactions between compensation effectors and the core clock network using the model eukaryote Neurospora crassa. Although research and training were significantly disrupted by the COVID-19 pandemic, the reverse genetic screen for mutants with defects in TC and NC was completed using the Neurospora knockout collection (Aim 1). The number of mutants with NC defects has more than doubled because of this work. The mass spectrometry experiments to identify TC-relevant substrates of Casein Kinase 1 and 2 are well underway, and publication is anticipated in 2022 (Aim 2). This Administrative Supplement will support additional months of professional development training lost to the COVID-19 pandemic as well as publication of a third first-author manuscript under this NRSA fellowship. Molecular mechanisms of circadian compensation are beginning to take shape due to research completed under this project. This and future independent investigator projects will resolve how the ~24-hour circadian period length is maintained in the face of different nutrient levels and temperatures in the environment, findings that have implications for humans with circadian sleep disorders, shift workers, continuous jet-lag, and other chronobiological conditions.