Initiation of DNA Replication at Cell Origins in Yeast

酵母细胞起源处 DNA 复制的起始

• 批准号: 8470651
• 负责人: BRUCE W. STILLMAN
• 金额: $ 67.62万
• 依托单位: COLD SPRING HARBOR LABORATORY
• 依托单位国家: 美国
• 财政年份: 1991
• 资助国家: 美国
• 起止时间: 1991-07-01 至 2016-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8470651
• 关键词: Address; Applications Grants; Area; Award; Biochemistry; Biological Models; Cells; Chromatin; Chromatin Modeling; Chromatin Structure; Chromosomes; Complex; Coupled; DNA; DNA Damage; DNA Replication Factor; DNA Sequence; DNA biosynthesis; DNA replication fork; DNA replication origin; Diagnosis; Disease; Epigenetic Process; Eukaryotic Cell; Funding; G1 Phase; Generations; Genome; Genome Stability; Genomic Instability; Germ Lines; Goals; Heart; Histones; Human; In Vitro; Inherited; Interruption; Laboratories; Lead; MCM2 gene; Maintenance; Meiosis; Mitosis; Mitotic; Mutation; Neoplastic Cell Transformation; Nucleosomes; Postdoctoral Fellow; Process; Protein Kinase; Proteins; Regulation; Replication Initiation; Research; Research Support; Resolution; Role; S Phase; Saccharomyces cerevisiae; Series; Signal Transduction; Sister Chromatid; Somatic Cell; System; Technology; Therapeutic Intervention; Time; Work; Yeasts; base; biological systems; cancer cell; career; chromatin assembly factor I; follow-up; histone modification; inhibitor/antagonist; origin recognition complex; reconstitution; response; segregation

DESCRIPTION (provided by applicant): The goal of this research is to determine the mechanism and regulation of the initiation of DNA replication in eukaryotic cells. It is clear that for maintenance of the integrity of the genome frm one cell generation to the next, DNA and its associated chromatin structures must be duplicated in a highly controlled and accurate manner. Interruption of these controls may promote genome instability and lead to neoplastic transformation in somatic cells or result in mutations in the germ line that can cause many different disorders. Moreover, the DNA replication proteins represent tangible targets for therapeutic intervention and diagnosis of proliferation of cancer cells, and other proliferative disorders. The initiator protein (ORC) cooperates with a series of DNA replication proteins, including Cdc6, Cdt1 and the MCM2-7 hexamer to establish at origins of DNA replication a pre-Replicative Complex (pre-RC) that facilitates later initiation of DNA synthesis at each origin. Recent progress has enabled the assembly of the pre-RC in vitro with purified proteins. The proposed research in this application will investigate, using the yeast S. cerevisiae, how the initiation of DNA replication occurs following pre-RC assembly and how this process is regulated by the Cdc7-Dbf4 (DDK) protein kinase and by an intrinsic inhibitor of initiation of DNA replication within the Mcm4 subunit of the MCM2-7 complex. The proposed research will also investigate how the core histones within nucleosomes, the fundamental structural unit of chromatin in eukaryotic cells, are disrupted during DNA replication and transferred to the leading and lagging strands of the newly synthesized DNA.

描述（由申请人提供）： 本研究的目的是确定真核细胞中DNA复制起始的机制和调控。很明显，为了保持基因组从一代到下一代的完整性，DNA及其相关的染色质结构必须以高度受控和准确的方式复制。这些控制的中断可能会促进基因组不稳定性，并导致体细胞中的肿瘤转化或导致生殖系中的突变，从而导致许多不同的疾病。此外，DNA复制蛋白代表了用于治疗干预和诊断癌细胞增殖和其他增殖性病症的有形靶标。起始蛋白（ORC）与一系列DNA复制蛋白（包括Cdc 6、Cdt 1和MCM 2 -7六聚体）合作，在DNA复制起点处建立前复制复合物（pre-Replicative Complex，pre-RC），促进随后在每个起点处启动DNA合成。最近的进展已经使得前RC在体外与纯化的蛋白质组装成为可能。本研究将利用酵母菌S.本发明的目的在于研究在酿酒酵母中的DNA复制的起始，在pre-RC组装之后如何发生DNA复制的起始，以及该过程如何被Cdc 7-Dbf 4（DDK）蛋白激酶和MCM 2 -7复合物的Mcm 4亚基内的DNA复制起始的内在抑制剂调节。拟议的研究还将调查核小体内的核心组蛋白，真核细胞染色质的基本结构单位，如何在DNA复制过程中被破坏，并转移到新合成的DNA的前导链和滞后链。