Project 1: Chromosome Inheritance

项目1：染色体遗传

• 批准号: 10092122
• 负责人: BRUCE W. STILLMAN
• 金额: $ 65.76万
• 依托单位: COLD SPRING HARBOR LABORATORY
• 依托单位国家: 美国
• 财政年份: 1997
• 资助国家: 美国
• 起止时间: 1997-02-10 至 2023-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10092122
• 关键词: Acute Myelocytic Leukemia; Adult; Antisense Oligonucleotides; Area; Binding; Binding Proteins; Breast; Breast Cancer Cell; Breast Cancer Model; CCNE1 gene; CDC6 gene; CDK2 gene; Carcinoma; Cell Cycle; Cell Proliferation; Cell division; Cell physiology; Cells; Centrioles; Centromere; Centrosome; Chromosome Segregation; Chromosome abnormality; Chromosomes; Colon; Complex; Cyclin A; Cyclin-Dependent Kinases; Cytokinesis; DNA Replication Timing; DNA biosynthesis; DNA replication fork; Data; Defect; Dependence; Disease; E2F transcription factors; Ensure; Epithelial; Gene Expression; Genes; Genome; Genome Stability; Goals; Hematopoietic; Heterochromatin; Human; Human Genome; In Vitro; Inherited; Kinetochores; Laboratories; Malignant Neoplasms; Mediating; Messenger RNA; Metaphase Plate; Microtubules; Mitosis; Mitotic; Mus; Mutation; NCI Center for Cancer Research; Nature; Normal Cell; ORC1L gene; Phosphotransferases; Pre-Replication Complex; Process; Proliferating; Prophase; Proteins; RNA Binding; RNA Helicase; Regulation; Replication Initiation; Replication Origin; Repression; Research; Retinoblastoma Protein; Role; S Phase; Series; Simian virus 40; Site; Testing; Time; Tissues; Tumor Suppressor Proteins; Untranslated RNA; Yeasts; acute myeloid leukemia cell; base; cancer cell; experimental study; falls; genetic information; histone methyltransferase; insight; leukemia; malignant breast neoplasm; mouse model; neoplastic cell; novel therapeutic intervention; origin recognition complex; reconstitution; recruit; segregation; therapeutic target; tumor

PROJECT SUMMARY- PROJECT 1 Cancer cells display uncontrolled inheritance of chromosomes, including errors in DNA replication and mitosis. These errors cause and propagate mutations and chromosomal abnormalities that further enhance cancer. Project 1 has been a leader in studying the mechanisms and control of inheritance of the human genome and has identified many of the key proteins that are involved in DNA synthesis at the replication fork and other proteins that are involved in the initiation of DNA replication. In the proposed studies, Project 1 will continue focusing on the mechanism and regulation of the initiation of DNA replication. Project 1 has discovered that certain initiation proteins are involved in many aspects of the cell division cycle, including centrosome duplication, centromere function and cytokinesis. Recent results also show that some initiation proteins are intimately involved in the fundamental decision of whether newly born cells will commit to a new round of cell division or enter into a period of quiescence. The proposed research will fall into three areas. Specific Aim 1 will focus on the role of the Origin Recognition Complex (ORC) subunit ORC1 and its related protein CDC6 in regulation of the commitment to cell division by controlling the expression of E2F1-regulated genes, in addition to the gene encoding Cyclin E, in cooperation with the tumor suppressor protein RB and the histone methyltransferase SUV39H1. Aim 1 will also focus on how CDC6 cooperates with ORC, RB, SUV39H1 and Cyclin dependent kinases to promote initiation of DNA replication. Specific Aim 2 will continue to study the role of the ORC subunits ORC2 and ORC3 at centromeres. ORC2 interacts with the Spindle Assembly Checkpoint protein BUBR1 only when it is phosphorylated during mitosis and defects in the BUBR1 binding domain of ORC2 cause the persistence of chromosomes that fail to align at the metaphase plate. Aim 2 will determine how ORC2 binds to BUBR1 and controls access of BUBR1. How ORC2 and ORC3 are recruited to centromeres, and the role of ORC3 interaction with the HP1 heterochromatin protein in chromosome segregation will also be studied. Project 3 will also study E2f1-regulated control of cell proliferation in a subset of breast and colon epithelial cancers have an acquired dependence of the DEAD-box RNA helicase DDX5. DDX5 is also required for progression of Acute Myeloid Leukemia (AML). This Aim will investigate how DDX5 becomes essential in some adult cancers and AML, while dispensable in normal epithelial and normal hematopoietic cells. Both RNA and protein binding partners will be identified, comparing both DDX5-dependent (DDX5-D) and DDX5-independent (DDX5-I) breast cancer cells. Exploiting the differential dependence on DDX5, Anti-Sense Oligonucleotides (ASO) will be developed for studying the role of DDX5 in mouse models for breast cancer and AML.

项目摘要 - 项目1 癌细胞表现出不受控制的染色体遗传，包括DNA复制和有丝分裂的错误。 这些误差会导致并传播突变和染色体异常，从而进一步增强癌症。 项目1一直是研究人类基因组遗传机制和控制的领导者 已经确定了许多在复制叉和其他的DNA合成中涉及的关键蛋白 参与DNA复制的蛋白质。在拟议的研究中，项目1将继续 专注于DNA复制开始的机理和调节。项目1发现 某些起始蛋白参与细胞分裂周期的许多方面，包括中心体 重复，丝粒功能和细胞因子。最近的结果还表明，某些起始蛋白是 密切参与了新出生的细胞是否会致力于新的细胞的基本决定 分裂或进入静止期。拟议的研究将分为三个领域。具体目标1 将重点关注原点识别复合物（ORC）亚基ORC1及其相关蛋白Cdc6在 通过控制E2F1调节基因的表达来调节对细胞分裂的承诺，此外 与编码细胞周期蛋白E的基因，与肿瘤抑制蛋白RB和组蛋白合作 甲基转移酶SUV39H1。 AIM 1还将重点介绍CDC6如何与ORC，RB，SUV39H1和 细胞周期蛋白依赖性激酶以促进DNA复制的开始。具体目标2将继续研究角色 Centromeres的ORC亚基ORC2和ORC3。 ORC2与主轴组件检查点相互作用 仅当在有丝分裂过程中磷酸化并在BUBR1结合结构域中磷酸化时，蛋白质BUBR1才被磷酸化。 ORC2导致无法在中期板上对齐的染色体的持久性。 AIM 2将确定 ORC2与BUBR1结合并控制BUBR1的访问。如何招募ORC2和ORC3 Centromeres，以及ORC3与HP1异染色质蛋白在染色体中的作用 也将研究隔离。项目3还将研究子集中E2F1调节的细胞增殖的控制 乳腺癌和结肠上皮癌的依赖性对死盒RNA解旋酶DDX5的依赖性。 急性髓样白血病（AML）的进展也需要DDX5。这个目标将调查DDX5如何 在某些成年癌和A​​ML中成为必不可少的 造血细胞。将鉴定RNA和蛋白质结合伙伴，并比较ddx5依赖性 （DDX5-D）和DDX5非依赖性（DDX5-I）乳腺癌细胞。利用差异依赖性 将开发DDX5，抗义寡核苷酸（ASO）来研究DDX5在小鼠模型中的作用 用于乳腺癌和AML。