Studies on Eukaryotic Replication

真核复制研究

• 批准号: 7903851
• 负责人: Jerard Hurwitz
• 金额: $ 30.99万
• 依托单位: SLOAN-KETTERING INST CAN RESEARCH
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-08-31 至 2011-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7903851
• 关键词: Binding; Cell Cycle; Cell Death; Cells; Complex; Coupled; DNA; DNA Primase; DNA biosynthesis; DNA-Directed DNA Polymerase; Event; Goals; Grant; Hereditary Disease; Human; Individual; Investigation; Laboratories; Lead; Malignant Neoplasms; Phosphotransferases; Polymerase; Pre-Replication Complex; Process; Property; Proteins; Reaction; Research; Role; Work; cohesin; helicase; in vivo; novel; protein complex

DESCRIPTION (provided by applicant): The goal of our research is to identify and characterize the machinery involved in eukaryotic DNA replication. We focus on the isolation of the individual proteins and protein complexes required for DNA synthesis and define their role in this critical macromolecular process. All growing cells must duplicate their DNA accurately and perform this feat once and only once per cell cycle. Events that interfere with these requirements can lead to cell death or altered proliferation that result in cancers. At present, many steps involved in supporting replication and its control are either unknown or poorly characterized. We plan to extend our previous work on various key replication proteins and their complexes that support the activation of initiation complexes required for the elongation of DNA chains. For this purpose, we will study: a) the role of a complex, containing Cdc45, the MCM2-7 complex and GINS (CMG) which contains DNA helicase activity and may contribute important to the unwinding reaction at the replication fork; b) the interaction between GINS and the Pol 1-primase complex which markedly stimulates its DNA polymerase activity; c) the properties of cloned Pol5 and its interaction with PCNA required for its processive elongation of primed DNA templates. We have also observed functional interactions of GINS with Pol5 as well as Pol4. We plan to examine these effects further and evaluate whether the CMG complex does the same. Coupled replication and unwinding reactions will be carried out which may contribute to strand selection; d) We plan to explore the role of the Cdc7 kinase on the activity and formation of the CMG complex. In vivo, this kinase was shown to influence the interaction of Cdc45 with the MCM2-7 complex. We will further characterize a novel interaction recently detected between the Cdc7 kinase complex and the cohesin loader complex Scc2/Scc4. The requirements for this interaction and its consequences will be investigated. All cells must make identical copies of their DNA to survive. Our studies are directed at how these copies are made and regulated. We plan to isolate the proteins involved in copying DNA and determine how they function. Results from these studies will have a direct impact on genetic diseases and cancer.

描述（由申请人提供）：我们的研究目标是鉴定和表征真核生物DNA复制所涉及的机制。我们专注于分离DNA合成所需的单个蛋白质和蛋白质复合物，并确定它们在这一关键大分子过程中的作用。所有生长中的细胞都必须准确地复制它们的DNA，并且在每个细胞周期中只能复制一次。干扰这些需求的事件可能导致细胞死亡或增殖改变，从而导致癌症。目前，支持复制及其控制的许多步骤要么是未知的，要么是缺乏特征的。