Identification and Analysis of Circadian Clock-Controlled Genes

描述(由申请人提供)：几乎所有被适当检查的真核生物都被证明具有内源性时间控制和组织的能力，称为昼夜节律。负责产生节律的细胞机械统称为生物钟。一个健康的生物钟是身体和心理健康的基础。由于时钟对人类心理和生理过程的影响无处不在--从人类基本生理学的昼夜变化到工作/休息周期和睡眠中明显的节奏--理解时钟对于预防和治疗许多身体和精神疾病是基本的。 我们的具体目标是为了加深我们对时钟调节细胞行为的方式的理解，包括细胞生长，在某些情况下还包括癌症。具体目标#1建立在我们将细胞周期调节蛋白检查点蛋白2确定为脉孢子菌中时钟功能的条件调节器的基础上。我们将在哺乳动物细胞、培养和小鼠身上继续这项工作，以探索昼夜节律系统和细胞周期的相互联系。具体目标2建立在我们对脉孢菌非昼夜节律输出振荡器的持续分析的基础上，将其作为调节昼夜节律输出的重要手段。我们将确定输出振荡器的组件，并确定它们连接到昼夜节律周期的方式。具体目标3建立在我们在基因组学方面的坚实基础上。我们将对输出振荡器的成分进行过表达筛选和敲除筛选，并将使用微阵列来鉴定脉孢子菌中钟控和输出振荡器控制的基因。 我们的长期目标是用遗传学和生物化学的语言描述包括生物钟的反馈周期，这个周期是如何与环境同步的，以及反馈周期产生的时间信息如何被用来调节细胞和有机体的行为。这些项目是互补和相互丰富的，因为它们依赖于基因和分子技术来剖析并最终理解作为时间函数的细胞的组织。公共卫生相关性：几乎所有经过适当检查的真核生物都被证明具有内源性时间控制和组织的能力，称为昼夜节律，即生物钟的输出。一个健康的生物钟是身体和心理健康的基础。由于时钟对人类心理和生理过程的影响无处不在--从人类基本生理学的昼夜变化到工作/休息周期和睡眠中明显的节奏--理解时钟对于预防和治疗许多身体和精神疾病是基本的。

DESCRIPTION (provided by applicant): Virtually all eukaryotic organisms appropriately examined have been shown to possess the capacity for endogenous temporal control and organization known as a circadian rhythm. The cellular machinery responsible for generating rhythms is collectively known as the biological clock. A healthy circadian clock underlies both physical and mental health. Because of the ubiquity of its influence on human mental and physiological processes - from circadian changes in basic human physiology to the clear involvement of rhythms in work/rest cycles and sleep - understanding the clock is basic to prevention and treatment of many physical and mental illnesses. Our specific aims are designed to further our understanding of the means through which the clock regulates cell behavior, including cell growth and in some cases cancer. Specific Aim #1 builds upon our identification of the cell cycle regulator checkpoint kinase 2 as a conditional modifier of clock function in Neurospora. We will pursue this work in mammalian cells in culture and in mice to probe the interconnectedness of the circadian system and cell cycle. Specific Aim #2 builds upon our ongoing analysis of non-circadian output oscillators in Neurospora as an important means of mediating circadian output. We will identify components of output oscillators and determine the means through which they are connected to the circadian cycle. Specific Aim #3 builds upon our strong grounding in genomics. We will carry out an overexpression screen and a knockout screen for components of output oscillators and will use microarrays to identify the clock-controlled and output oscillator-controlled genes in Neurospora. Our long term goals are to describe, in the language of genetics and biochemistry, the feedback cycle comprising the circadian clock, how this cycle is synchronized with the environment, and how time information generated by the feedback cycle is used to regulate the behavior of cells and organisms. These projects are complementary and mutually enriching in that they rely on genetic and molecular techniques to dissect, and ultimately to understand, the organization of the cell as a function of time. PUBLIC HEALTH RELEVANCE: Virtually all eukaryotic organisms appropriately examined have been shown to possess the capacity for endogenous temporal control and organization known as a circadian rhythm, the output of a biological clock. A healthy circadian clock underlies both physical and mental health. Because of the ubiquity of its influence on human mental and physiological processes - from circadian changes in basic human physiology to the clear involvement of rhythms in work/rest cycles and sleep - understanding the clock is basic to prevention and treatment of many physical and mental illnesses.