Chromosome Inheritance

染色体遗传

• 批准号: 8234410
• 负责人: BRUCE W. STILLMAN
• 金额: $ 60.89万
• 依托单位: COLD SPRING HARBOR LABORATORY
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-01-01 至 2016-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8234410
• 关键词: Anaphase; Animal Model; Binding; Boxing; Breast; Breast Cancer Cell; CDC6 gene; CDK2 gene; Cancer Control; Cell Aging; Cell Cycle; Cell Death; Cell Proliferation; Cell Therapy; Cell physiology; Cells; Centrioles; Centromere; Centrosome; Characteristics; Chromosome Segregation; Chromosome Structures; Chromosomes; Complex; Cyclin A; Cyclin E; Cytokinesis; DNA; DNA biosynthesis; DNA replication origin; Defect; Drosophila genus; E2F Transcription Factor 1; E2F1 gene; Elements; Ensure; Environment; G1 Phase; Gene Expression; Genes; Genetic; Genome; Genome Stability; Goals; Heterochromatin; Human; Human Genome; In Vitro; Instruction; Kinetochores; Laboratories; Location; Maintenance; Malignant Neoplasms; Mammary Neoplasms; Metaphase; Methods; Microtubules; Mitosis; Mitotic; Mus; Mutation; NCI Center for Cancer Research; Nature; Normal Cell; Nuclear; Organism; Phase; Phosphorylation; Phosphotransferases; Play; Process; Proliferating; Prophase; Proteins; Proteomics; RNA Helicase; Replication Initiation; Research; Roche brand of trastuzumab; Role; S Phase; Signal Transduction; Simian virus 40; Site; Testing; Time; Tissues; Trastuzumab; Work; Yeasts; base; cancer cell; cancer therapy; chromatin protein; insight; malignant breast neoplasm; neoplastic cell; next generation; origin recognition complex; protein complex; reconstitution; research study; segregation; therapeutic target; tumor; tumor xenograft

PROJECT SUMMARY (See instructions): Cancer involves the induction of uncontrolled DNA replication and mitosis in cells, as well as processes that ensure cells evade cell death or senescence and survive in specific tissue micro-environments. 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 to focus on how the initiation of DNA replication is controlled in human cells and how this process goes awry in tumor cells. Specific Aim 1 will focus on how the origins of DNA replication are marked in chromosomes so that they can form pre-replicative complexes during exit from mitosis or during Gl phase, thereby enabling the initiation of DNA replication in S phase of the cell division cycle. Since the Origin Recognition Complex, particularly its largest subunit Orel is loaded onto chromosomes beginning in prophase of mitosis and Orel is the most stably bound chromatin protein, the locations within the human genome for Orel binding will be determined. In addition, proteins that dynamically interact with ORC during M and G1 phases will be determined. The cell cycle regulators Cyclin E-CDK2 and Cyclin A-CDK2, the former often over-active in breast cancer, are controlled by direct interactions with Orel and Cdc6 and how these interactions influence the initiation of DNA replication and centriole duplication in centrosomes will be investigated. Recent evidence has emerged that ORC subunits play a critical role at kinetochores that bind microtubule spindles for congression of chromosomes prior to their segregation. In Specific Aim 2, interactions between the Orc2 and Orc3 subunits of ORC and the Spindle Assembly Checkpoint kinase BubRI will be investigated, as will the role of the ORC subunits in maintenance of stable spindle attachment during the metaphase to anaphase transition. In Specific Aim 3, the control of DNA replication by the DEAD-box RNA helicase DDX5 and its interaction with the transcription factor E2F1 will be studied. DDX5 is amplified in the genome of cells in 25% of human breast cancers and it is these cells that display selective sensitivity to inhibition of DDX5 protein levels. How DDX5 influences E2F1-driven expression of DNA replication genes in the Gl phase of the cell cycle will be investigated. Project 1 will also investigate how tumor cells with amplified copy number of DDX5 become addicted to its continued expression, in contrast to normal, non-tumor cells and many other cancer cells. Finally, the additive effects on inhibition of tumors cell proliferation with the combination of DDX5 depletion and Trastuzumab (Herceptin) treatment will be examined.

项目摘要（请参阅说明）： 癌症涉及细胞中不受控制的DNA复制和有丝分裂的诱导，以及确保细胞逃避细胞死亡或衰老并在特定组织微环境中存活的过程。项目1一直是研究人类基因组遗传的机制和控制的领导 参与DNA复制的蛋白质。在拟议的研究中。项目1将继续关注人类细胞中DNA复制的启动以及该过程如何在肿瘤细胞中出现问题。具体的目标1将重点介绍DNA复制的起源如何在染色体中标记，以便它们可以在出口脱离有丝分裂或GL期间形成复制的复合物，从而使 在细胞分裂周期的S相中，DNA复制的启动。由于起源识别复合物，特别是其最大的亚基Orel被加载到有丝分裂预言的染色体上，而Orel是最稳定的染色质蛋白，因此将确定人类基因组内的OREL结合中的位置。另外，将确定在M和G1相过程中与ORC动态相互作用的蛋白质。细胞周期调节剂Cyclin E-CDK2和细胞周期蛋白A-CDK2（前者通常在乳腺癌中过度活跃）受到与Orel和Cdc6的直接相互作用的控制，这些相互作用如何影响DNA复制和中心体中心体重复的启动。最近有证据表明，兽人亚基在隔离之前结合微管纺锤体的动力学上起着至关重要的作用。在特定的目标2中，将研究ORC的ORC2和ORC3亚基与纺锤体组件检查点激酶BUBRI之间的相互作用，ORC亚基在中期对过后期过渡过程中稳定纺锤体附着的稳定纺锤体附件中的作用也将进行。在特定的目标3中，将研究由死盒RNA解旋酶DDX5对DNA复制的控制及其与转录因子E2F1的相互作用。 DDX5在25％的人乳腺癌中的细胞基因组中扩增，正是这些细胞对抑制DDX5蛋白水平的抑制作用表现出选择性敏感性。 DDX5如何影响细胞周期GL相中DNA复制基因的E2F1驱动的表达。项目1还将调查与正常，非肿瘤细胞和许多其他癌细胞相比，DDX5的扩增拷贝数的肿瘤细胞如何上瘾。最后，将检查对抑制肿瘤细胞增殖与DDX5耗竭和曲妥珠单抗（赫赛汀）治疗的添加作用。