Inhibition of DNA re-replication by cyclin/CDK and GSK-3 kinase in S. cerevisiae

酿酒酵母中细胞周期蛋白/CDK 和 GSK-3 激酶对 DNA 再复制的抑制

基本信息

批准号：
8473647
负责人：
Amy E Ikui
金额：
$ 11.78万
依托单位：
BROOKLYN COLLEGE
依托单位国家：
美国
项目类别：
财政年份：
2013
资助国家：
美国
起止时间：
2013-07-01 至 2017-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8473647
关键词：
Address Amino Acid Sequence Bacteria Biochemical Biological Assay Cell Cycle Cell physiology Chromosomal Instability Chromosomes Complex Consensus Cyclin-Dependent Kinases Cyclins DNA DNA Replication Factor DNA biosynthesis Development Ensure Eukaryota Eukaryotic Cell Failure Future Genetic Epistasis Genome Genomic Instability Glycogen Synthase Kinase 3 Goals Human In Vitro Investigation Knowledge Lead Licensing Life Link Mediating Molecular Mutation N-terminal Organism Outcomes Research Phosphorylation Phosphotransferases Prevention Protein Kinase Proteins Recombinants Replication-Associated Process Role Saccharomyces cerevisiae Saccharomycetales Site System Testing Time Ubiquitination Yeasts base cancer therapy college design inhibitor/antagonist innovation mutant novel public health relevance research study therapeutic target tumorigenesis ubiquitin-protein ligase yeast genetics

项目摘要

DESCRIPTION (provided by applicant): Eukaryotic cells duplicate their chromosomes. Errors in the DNA replication control could lead to chromosome instability. Our long-term goal is to understand the molecular mechanism of DNA replication control in eukaryotes. Cdc6p is a key DNA replication component from yeast to humans. Our preliminary results showed that yeast GSK-3 kinase (Mck1p) targets DNA replication factor Cdc6p for degradation, which in turn inhibits DNA re-replication. It is known that N-terminal region of Cdc6 is phosphorylated by Cyclin/CDK complex for its degradation after the origin is licensed. We will study the novel Cdc6p degradation mechanism controlled by Mck1p kinase to inhibit DNA re- replication. In this proposal, three aims will test my hypothesis if a sequential phosphorylation of Cdc6p by Cdk1 and GSK3 controls the timing of Cdc6p degradation. Aim1 will test if Cdc6p is directly phosphorylated by Mck1 kinase, and Mck1-depedent Cdc6 degradation is mediated through SCFCDC4 complex. Aim2 will test the possibility that Mck1p targets Cdc6p in a distinct mechanism from CDK. Aim3 will test if the phosphorylation of Cdc6 by CDK is a pre-requisite for phosphorylation by Mck1p. Cdc6p and GSK-3 kinase are conserved from yeast to humans. The results obtained from this study can be applied to understand the molecular mechanism of Cdc6 degradation by GSK-3 kinase in higher eukaryotes. The accurate DNA replication control is a critical step to avoid chromosome instability, and it is important to understand since the chromosome instability is a hallmark of tumorigenesis.

描述（由申请人提供）：真核细胞复制其染色体。 DNA 复制控制中的错误可能导致染色体不稳定。我们的长期目标是了解真核生物中 DNA 复制控制的分子机制。 Cdc6p 是从酵母到人类的关键 DNA 复制组件。我们的初步结果表明，酵母 GSK-3 激酶 (Mck1p) 以 DNA 复制因子 Cdc6p 为目标进行降解，从而抑制 DNA 重新复制。已知Cdc6的N端区域在来源获得许可后被Cyclin/CDK复合物磷酸化并降解。我们将研究由Mck1p激酶控制的新型Cdc6p降解机制来抑制DNA复制。在这个提案中，三个目标将检验我的假设：Cdk1 和 GSK3 对 Cdc6p 的连续磷酸化是否控制了 Cdc6p 降解的时间。 Aim1 将测试 Cdc6p 是否直接被 Mck1 激酶磷酸化，以及 Mck1 依赖性的 Cdc6 降解是通过 SCFCDC4 复合物介导的。 Aim2 将测试 Mck1p 以与 CDK 不同的机制靶向 Cdc6p 的可能性。 Aim3 将测试 CDK 对 Cdc6 的磷酸化是否是 Mck1p 磷酸化的先决条件。 Cdc6p 和 GSK-3 激酶从酵母到人类都是保守的。本研究获得的结果可用于了解高等真核生物中GSK-3激酶降解Cdc6的分子机制。准确的DNA复制控制是避免染色体不稳定的关键步骤，并且理解这一点很重要，因为染色体不稳定是肿瘤发生的标志。