Anaphase Promoting Complex-mediated Proteolysis

后期促进复合物介导的蛋白水解

• 批准号: 7540389
• 负责人: MARK J SOLOMON
• 金额: $ 34.13万
• 依托单位: YALE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-01-01 至 2010-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7540389
• 关键词: Adenomatous Polyposis Coli Protein; Binding; Boxing; Cell Cycle; Cell Proliferation; Cell division; Cells; Chromosomes; Complex; Cyclins; Disease; Event; Genetic Screening; Goals; Human; Immunoblotting; Malignant Neoplasms; Mediating; Mitosis; Mitotic spindle; Modeling; Molecular Conformation; Normal Cell; Pathway interactions; Phase; Phosphorylation; Process; Proteins; Proteolysis; Research Personnel; Role; Saccharomycetales; System; Testing; Ubiquitin; Ubiquitination; Work; Yeasts; anaphase-promoting complex; insight; man; novel; overexpression; prevent; programs; research study

A key event in the progression through and exit from mitosis is the ubiquitin-dependent degradation of cyclins and other key proteins by the Anaphase Promoting Complex (ARC; also called the cyclosome).ARC substrates contain degradation motifs such as the Destruction Box and the KEN Box that are required for their ubiquitination and for their binding to the ARC activators, Cdc20 and Cdh1. Both motifs are required for efficient degradation, but either one alone suffices for binding to Cdhlp. We have proposed a pathway for the assembly of the APC-Cdhip-substrate complex in which APC-free Cdhlp first binds a substrate. Engagement of the substrate Destruction Box by Cdh1p stimulates the binding of Cdh1p to the ARC, presumably via a conformation change in Cdhlp. The current studies are aimed at furthering our understanding of APC-mediated protedlysis in budding yeast. In particular, we wish to identify novel ARC substrates, to understand how APC-Cdh1p/Cdc20p-substrate complexes are assembled and disassembled, and to understand how the spindle assembly checkpoint makes use of a KEN box to turn off APC-mediated proteolysis when chromosomes are not properly attached to the mitotic spindle. To achieve these goals, I propose the following Specific Aims: 1) To Identify Novel ARC Substrates. 2) To Assess the Generality of the APC-Cdhip-Substrate Assembly Pathway. 3) To Determine How D-Box Engagement Stimulates Cdh1p Binding to the ARC: 4) To Determine the Roles of Conserved Cdhlp Motifs and of Phosphcrylation in APC-Cdh1p-Substrate Assembly. 5) To Determine Whether Disassembly of the APC-Cdh1p-Substrate Complex is an Active Process. 6) To Determine the Function of the MadSp KEN Box in the Spindle Assembly Checkpoint. Degradation of key proteins is essential for cell division. Understanding this degradation is thus important for understanding normal cell proliferation, and how this process goes awry in disease states, particularly cancers.

通过有丝分裂进展和退出的关键事件是依赖泛素的降解 促进复合物（弧；也称为旋风体）的细胞周期蛋白和其他关键蛋白。arc 底物包含降解图案，例如破坏框和ken框 它们的泛素化及其与电弧激活剂CDC20和CDH1的结合。两个图案都是必需的 为了有效的降解，但任何一个人都足以与CDHLP结合。我们提出了一条途径 对于APC-CDHIP-SUBSTRATE复合物的组装，其中无APC CDHLP首先结合底物。 CDH1P对基板破坏框的参与刺激CDH1P与ARC的结合， 大概是通过CDHLP的构象变化。 当前的研究旨在进一步了解APC介导的普通性呈萌芽状态 酵母。特别是，我们希望识别新颖的弧底物，以了解 APC-CDH1P/CDC20P-SUBSTRATE络合物组装和拆卸，并了解如何了解 主轴组件检查站使用KEN盒子关闭APC介导的蛋白水解 染色体未正确连接到有丝分裂主轴上。 为了实现这些目标，我提出以下具体目标： 1）识别新型弧底物。 2）评估APC-CDHIP-SUBSTRATE组装途径的一般性。 3）确定d-box参与如何刺激CDH1P与弧的结合： 4）确定保守的CDHLP基序的作用和APC-CDH1P-SUBSTRATE中磷酸化的作用 集会。 5）确定APC-CDH1P-SUBSTRATE复合物的拆卸是一个活动过程。 6）确定主轴组件检查点中MADSP KEN框的功能。 关键蛋白的降解对于细胞分裂至关重要。因此，了解这种退化是 对于理解正常细胞增殖以及该过程如何在疾病状态下出现问题很重要， 特别是癌症。