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 癌细胞显示染色体的不受控制的遗传，包括DNA复制和有丝分裂中的错误。 这些错误导致并传播突变和染色体异常，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， 已经确定了许多关键蛋白质，这些蛋白质在复制叉和其他 参与DNA复制起始的蛋白质。在拟议的研究中，项目1将继续 专注于DNA复制起始的机制和调节。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 复制、着丝粒功能和胞质分裂。最近的研究结果还表明，一些起始蛋白是 它与新生细胞是否会进入新一轮细胞分化的基本决定密切相关， 分裂或进入一个安静的时期。拟议的研究将分为三个领域。具体目标1 将重点关注起源识别复合物（ORC）亚基ORC 1及其相关蛋白CDC 6在 通过控制E2 F1调节基因的表达来调节细胞分裂的承诺，此外， 与肿瘤抑制蛋白RB和组蛋白 甲基转移酶SUV 39 H1。目标1还将重点介绍CDC 6如何与ORC、RB、SUV 39 H1和 促进DNA复制起始的细胞周期蛋白依赖性激酶。具体目标2将继续研究 ORC亚基ORC 2和ORC 3在着丝粒上。ORC 2与主轴组件检查点相互作用 蛋白质BUBR 1只有当它在有丝分裂过程中被磷酸化和缺陷的BUBR 1结合结构域， ORC 2导致染色体在中期板上无法对齐的持久性。目标2将决定 ORC 2如何绑定到BUBR 1并控制BUBR 1的访问。ORC 2和ORC 3是如何被招募的 着丝粒，以及ORC 3与染色体中HP 1异染色质蛋白相互作用的作用 此外，亦会研究种族隔离的问题。项目3还将研究E2 f1调节的细胞增殖控制在一个子集 乳腺癌和结肠上皮癌患者对DEAD盒RNA解旋酶DDX 5具有获得性依赖性。 DDX 5也是急性髓系白血病（AML）进展所必需的。本文将探讨DDX 5 在一些成人癌症和AML中是必需的，而在正常上皮和正常 造血细胞将鉴定RNA和蛋白质结合配偶体，比较DDX 5依赖性 在一些实施方案中，本发明涉及DDX 5-D和DDX 5非依赖性（DDX 5-I）乳腺癌细胞。利用微分依赖于 DDX 5，反义寡核苷酸（阿索）将用于研究DDX 5在小鼠模型中的作用 治疗乳腺癌和急性髓细胞白血病