The role of clock protein phosphorylation and degradation in circadian biology

时钟蛋白磷酸化和降解在昼夜节律生物学中的作用

• 批准号: 7447989
• 负责人: JOANNA Chungyen CHIU
• 金额: $ 8.71万
• 依托单位: RUTGERS, THE STATE UNIV OF N.J.
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-05-01 至 2010-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7447989
• 关键词: 26S proteasome; Affect; Affinity Chromatography; Alcohol dependence; Animal Model; Animals; Automobile Driving; Behavioral; Binding; Biology; CHK gene; CSNK2A1 gene; Cell Nucleus; Choline Kinase; Chronic; Circadian Rhythms; Classification; Clock protein; Condition; Cytoplasm; Daily; Defect; Degradation Pathway; Disease; Docking; Drosophila genus; Drosophila melanogaster; Elderly; Equilibrium; Event; F Box Domain; F-Box Proteins; Feedback; Goals; Homologous Gene; Human; Institution; Link; Lysine; Malignant Neoplasms; Mammals; Maps; Mass Spectrum Analysis; Mediating; Mentors; Metabolic syndrome; Metabolism; Modification; Molecular; Monitor; Mutagenesis; Mutation; Numbers; Output; Pathway interactions; Pharmaceutical Preparations; Phase; Phospho-Specific Antibodies; Phosphoric Monoester Hydrolases; Phosphorylation; Phosphorylation Site; Phosphotransferases; Physiological; Physiological Processes; Play; Predisposition; Protein Kinase; Protein phosphatase; Proteins; Public Health; Regulation; Research; Research Personnel; Role; Role playing therapy; Scientist; Seasonal Affective Disorder; Site; Sleep Disorders; System; Time; Ubiquitin; base; casein kinase; circadian pacemaker; day; genetic manipulation; in vitro Assay; in vivo; inhibitor/antagonist; insight; mRNA Expression; multicatalytic endopeptidase complex; novel; programs; protein function; protein protein interaction; tissue culture

DESCRIPTION (provided by applicant): Project Summary: The overall goal of this proposal is to better understand the contribution of clock protein phosphorylation and degradation pathways in regulating circadian rhythms by focusing on PERIOD (PER), the central clock protein. In animal clocks, PER undergoes daily oscillations in levels and phosphorylation states that are regulated by DOUBLETIME (DBT in Drosophila, CKIe in mammals), CKII, and phosphatase 2A. Hyperphosphorylated PER proteins are eventually targeted to the 26S proteasome by the F-box protein SLIMB (a homolog of B-TrCP in mammals). My immediate goal is to complete an additional 2 years of mentored research in Dr. Isaac Edery's lab. My long term goal is to become a tenure-tracked independent scientist at an academic institution, and contribute to a better understanding of how circadian oscillations are generated and maintained, and how they control the timing of output physiological processes. During the mentored phase, I plan to examine the mechanisms by which different phosphorylation events, identified from a mass spectral analysis of DBT-phosphorylated PER proteins, regulate PER function, subcellular localization, and interaction with other clock proteins. In addition, to better understand the SLIMB-mediated PER degradation pathway, and to build on my studies on PER-SLIMB interaction, I plan to dissect the steps of SLIMB-mediated PER degradation pathway following PER-SLIMB interaction. Upon transition into the independence phase, I propose to examine the phosphorylation profile of PER proteins isolated under other physiologically relevant conditions by mass spectral analysis, and examine the functional significance of the identified phosphosites in vivo. I believe this systematic approach will yield significant insights into the regulation of PER function by phosphorylation. Finally, I propose to use tandem affinity purification to isolate novel clock components and protein interactions that regulate PER phosphorylation, metabolism, and function. Relevance: The study of clock protein phosphorylation and metabolism has significant implications to public health. Mutations altering human PER2 phosphorylation or CKI kinase activity are linked to familial sleep disorders in humans. Taking a wider perspective, defects in circadian clock proteins have been implicated in other human disorders, including chronic sleep problems in the elderly, seasonal affective disorders, metabolic syndromes, and susceptibility to drug and alcohol addiction, and cancer.

描述（由申请人提供）： 项目摘要：该提案的总体目标是通过关注中央时钟蛋白 PERIOD (PER)，更好地了解时钟蛋白磷酸化和降解途径在调节昼夜节律中的贡献。在动物时钟中，PER 的水平和磷酸化状态每天都会发生波动，受 DOUBLETIME（果蝇中的 DBT，哺乳动物中的 CKIe）、CKII 和磷酸酶 2A 调节。过度磷酸化的 PER 蛋白最终通过 F-box 蛋白 SLIMB（哺乳动物中 B-TrCP 的同源物）靶向 26S 蛋白酶体。我的近期目标是在 Isaac Edery 博士的实验室完成另外 2 年的指导研究。我的长期目标是成为学术机构的终身教授独立科学家，并有助于更好地理解昼夜节律振荡是如何产生和维持的，以及它们如何控制输出生理过程的时间。在指导阶段，我计划研究通过 DBT 磷酸化 PER 蛋白的质谱分析确定的不同磷酸化事件调节 PER 功能、亚细胞定位以及与其他时钟蛋白相互作用的机制。此外，为了更好地理解 SLIMB 介导的 PER 降解途径，并以我对 PER-SLIMB 相互作用的研究为基础，我计划剖析 PER-SLIMB 相互作用后 SLIMB 介导的 PER 降解途径的步骤。过渡到独立阶段后，我建议通过质谱分析检查在其他生理相关条件下分离的 PER 蛋白的磷酸化谱，并检查已识别的磷酸位点在体内的功能意义。我相信这种系统方法将对磷酸化对 PER 功能的调节产生重要的见解。最后，我建议使用串联亲和纯化来分离调节 PER 磷酸化、代谢和功能的新时钟成分和蛋白质相互作用。相关性：时钟蛋白磷酸化和代谢的研究对公共健康具有重要意义。改变人类 PER2 磷酸化或 CKI 激酶活性的突变与人类家族性睡眠障碍有关。从更广泛的角度来看，生物钟蛋白的缺陷与其他人类疾病有关，包括老年人的慢性睡眠问题、季节性情感障碍、代谢综合征、对药物和酒精成瘾的易感性以及癌症。