Context-specific Functions of CDK8

CDK8 的上下文特定功能

• 批准号: 8824931
• 负责人: Jun-yuan Ji
• 金额: $ 30.64万
• 依托单位: TEXAS A&M UNIVERSITY HEALTH SCIENCE CTR
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-02-25 至 2016-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8824931
• 关键词: Address; Animal Model; Binding Proteins; Biological; Cardiovascular Diseases; Colorectal Cancer; Complex; Cues; Cyclin-Dependent Kinases; DNA Binding; Data; Defect; Development; Diabetes Mellitus; Disease; Drosophila genus; Drosophila melanogaster; Ecdysone; Eukaryota; Fatty acid glycerol esters; Feedback; Gene Expression; Gene Targeting; Genes; Genetic Transcription; Goals; Growth; Health; Homeostasis; Homologous Gene; Human; Insulin; Insulin Signaling Pathway; Larva; Link; Lipids; Malignant Neoplasms; Malignant neoplasm of liver; Mediating; Mediator of activation protein; Metabolism; Modeling; Molecular; Mutate; Nuclear Receptors; Nutrient; Obesity; Organism; Pathway interactions; Phenotype; Phosphorylation; Phosphotransferases; Physiological; Play; Proteins; RNA Polymerase II; RXR; Receptor Signaling; Recruitment Activity; Regulation; Regulatory Element; Research; Risk; Role; Signal Pathway; Signal Transduction; Staging; Sterols; Threonine; Time; Trans-Activators; Transcription Coactivator; Transcriptional Regulation; Up-Regulation; Work; cancer therapy; cancer type; cyclin C; ecdysteroid receptor; in vivo; insulin signaling; leukemia; lipid biosynthesis; lipid metabolism; loss of function mutation; melanoma; mutant; novel; programs; protein function; response; steroid hormone receptor; transcription factor; tumorigenesis

DESCRIPTION (provided by applicant): The highly conserved co-factor Mediator Complex, comprising up to 30 subunits, serves as a molecular bridge between transcriptional activators and the core transcription machinery, and is thought to be required for most RNA polymerase II-dependent transcription. As the only known enzymatic subunit of Mediator complex, CDK8 (Cyclin-Dependent Kinase 8) can either positively or negatively regulate transcription. However, whether CDK8 acts as an activator or repressor at every gene, or whether it acts differently at different classes of genes, is a key unexplored field that would dramatically refine our view of how Mediator facilitates transcription. Furthermore, understanding the function and regulation of CDK8 is critical to elucidate how dysregulation of both CDK8 and its regulatory partner CycC (Cyclin C) contributes to a variety of human cancers. Depletion of CDK8 effectively blocks the growth of melanoma and colorectal cancer, which underscores the centrality of CDK8 in gene expression and demonstrates why CDK8 is considered an attractive and promising target for cancer treatment. Currently, how dysregulation of CDK8-CycC contributes to tumorigenesis is poorly understood. To determine the normal and dysregulated functions of these proteins, it is essential to identify the downstream effectors and upstream regulators of CDK8-CycC. Analyses of function and regulation of CDK8 have been hampered by the lack of phenotypes for CDK8 activity in multicellular organisms. We have solved this challenge by using Drosophila, which provides unparalleled sophistication in manipulating CDK8 activity in vivo. We have recently identified two important downstream transactivators of CDK8, SREBP (Sterol Regulatory Element-Binding Protein) and the EcR (Ecdysteroid Receptor), which play essential roles in regulating lipogenesis and developmental maturation, respectively. Our analyses suggest that CDK8 inhibits SREBP-dependent transcription but activates EcR-activated gene expression. Thus, the objective of this proposal is to determine how CDK8 plays fundamentally different roles in modulating the activities of SREBP and EcR. Specifically, we will elucidate how CDK8 regulates SREBP- and EcR-mediated transcription by determining the molecular mechanism underlying the interactions between CDK8 and both SREBP and EcR, and the effects of SREBP and EcR phosphorylation by CDK8 in vivo. We will also examine the physiological regulation of CDK8-CycC by the insulin-signaling pathway and analyze the role of CDK8 in coordinating lipid homeostasis and developmental timing. This study will significantly advance our understanding of how CDK8 plays distinct roles in lipogenesis and development. Because CDK8 and CycC are well conserved in eukaryotes, the molecular mechanisms that we identify in Drosophila are highly likely to provide working models in human studies. The importance of this project is highlighted by the fact that dysregulation of lipid homeostasis durin development is closely linked to diseases such as diabetes, obesity, cardiovascular diseases, and certain types of cancers.

描述（由申请人提供）：高度保守的辅因子介体复合物，包含多达30个亚基，作为转录激活因子和核心转录机制之间的分子桥梁，被认为是大多数RNA聚合酶II依赖性转录所需的。CDK 8（Cyclin-Dependent Kinase 8）是介体复合物中唯一已知的酶亚基，对转录有正或负调控作用。然而，CDK 8是否在每个基因中充当激活剂或抑制剂，或者它是否在不同类别的基因中起不同的作用，是一个关键的未探索领域，这将极大地改善我们对介体如何促进转录的看法。此外，了解CDK 8的功能和调控对于阐明CDK 8及其调控伙伴CycC（Cyclin C）的失调如何导致各种人类癌症至关重要。CDK 8的消耗有效地阻止了黑色素瘤和结直肠癌的生长，这强调了CDK 8在基因表达中的中心地位，并证明了为什么CDK 8被认为是一个有吸引力和有前途的癌症治疗靶点。目前，对CDK 8-CycC的失调如何促进肿瘤发生知之甚少。为了确定这些蛋白质的正常和失调功能，必须确定CDK 8-CycC的下游效应子和上游调节子。CDK 8的功能和调节的分析受到了阻碍，缺乏CDK 8活性在多细胞生物体中的表型。我们已经通过使用果蝇解决了这一挑战，果蝇在体内操纵CDK 8活性方面提供了无与伦比的复杂性。我们最近发现了CDK 8的两个重要下游反式激活因子：SREBP（固醇调节元件结合蛋白）和EcR（蜕皮类固醇受体），它们分别在调节脂肪生成和发育成熟中发挥重要作用。我们的分析表明，CDK 8抑制SREBP依赖的转录，但激活ECR激活的基因表达。因此，本提案的目的是确定CDK 8如何在调节SREBP和EcR的活动中发挥根本不同的作用。具体来说，我们将阐明CDK 8如何调节SREBP和EcR介导的转录，通过确定CDK 8和SREBP和EcR之间的相互作用的分子机制，以及SREBP和EcR磷酸化的CDK 8在体内的影响。我们还将研究胰岛素信号通路对CDK 8-CycC的生理调节，并分析CDK 8在协调脂质稳态和发育时序中的作用。这项研究将大大推进我们对CDK 8如何在脂肪生成和发育中发挥独特作用的理解。由于CDK 8和CycC在真核生物中非常保守，因此我们在果蝇中鉴定的分子机制极有可能为人类研究提供工作模型。这个项目的重要性是突出的事实，脂质稳态失调在发展过程中密切相关的疾病，如糖尿病，肥胖症，心血管疾病和某些类型的癌症。