Chromosome Inheritance

染色体遗传

基本信息

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
项目类别：
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

项目摘要

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（参见说明）：癌症涉及诱导细胞内不受控制的DNA复制和有丝分裂，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生存的过程。项目 1 一直是研究人类基因组遗传机制和控制的领导者，并已鉴定出许多参与复制叉和其他部位 DNA 合成的关键蛋白质。参与 DNA 复制起始的蛋白质。在拟议的研究中。项目 1 将继续关注人类细胞中如何控制 DNA 复制的启动以及肿瘤细胞中这一过程如何出错。具体目标 1 将重点关注如何在染色体中标记 DNA 复制起点，以便它们能够在有丝分裂退出期间或 G1 期期间形成复制前复合物，从而使 DNA复制在细胞分裂周期的S期开始。由于起源识别复合体，特别是其最大的亚基 Orel 在有丝分裂前期开始被加载到染色体上，并且 Orel 是最稳定结合的染色质蛋白，因此将确定人类基因组中 Orel 结合的位置。此外，还将确定在 M 期和 G1 期与 ORC 动态相互作用的蛋白质。细胞周期调节因子 Cyclin E-CDK2 和 Cyclin A-CDK2（前者在乳腺癌中经常过度活跃）通过与 Orel 和 Cdc6 的直接相互作用来控制，以及这些相互作用如何影响中心体中 DNA 复制和中心粒复制的启动。调查了。最近的证据表明，ORC 亚基在动粒中发挥着关键作用，动粒在染色体分离之前结合微管纺锤体以进行染色体的聚集。在具体目标 2 中，将研究 ORC 的 Orc2 和 Orc3 亚基与纺锤体组装检查点激酶 BubRI 之间的相互作用，以及 ORC 亚基在中期到后期过渡期间维持纺锤体稳定附着中的作用。在具体目标 3 中，将研究 DEAD-box 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% 的人类乳腺癌细胞基因组中扩增，正是这些细胞对 DDX5 蛋白水平的抑制表现出选择性敏感性。将研究DDX5如何影响细胞周期G1期中E2F1驱动的DNA复制基因的表达。项目 1 还将研究与正常非肿瘤细胞和许多其他癌细胞相比，DDX5 拷贝数扩增的肿瘤细胞如何对其持续表达上瘾。最后，将检查 DDX5 去除和曲妥珠单抗（赫赛汀）治疗相结合对抑制肿瘤细胞增殖的累加效应。