DNA elements linking regulation of replication timing to oncogene function

将复制时间调控与癌基因功能联系起来的 DNA 元件

• 批准号: 8398254
• 负责人: Benjamin Daniel Pope
• 金额: $ 3.55万
• 依托单位: FLORIDA STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-08-07 至 2014-08-06
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8398254
• 关键词: Address; Cancer Etiology; Cell Nucleus; Cells; Characteristics; Chromatin; Chromosomes; DNA; DNA Replication Timing; DNA Sequence Rearrangement; Data; Development; Drug Formulations; Elements; Genetic; Genetic Transcription; Genome; Genomics; Goals; Indium; Individual; Knowledge; Lead; Link; Malignant Glioma; Malignant Neoplasms; Measures; Mission; National Cancer Institute; Nature; Oncogenes; Oncogenic; Outcome; Patients; Property; Public Health; Regulation; Regulatory Element; Replicon; Research; Role; Testing; Time; Work; cancer cell; cell growth regulation; cell type; gene function; insight; pleiotrophin; prevent; programs; promoter; relating to nervous system; research study; tumorigenesis

DESCRIPTION (provided by applicant): DNA replication timing and other coordinately regulated properties of chromatin organization are disrupted at oncogenic loci in cancer. Pleiotrophin (PTN) is an oncogene expressed in all malignant gliomas and is transcriptionally regulated during neural differentiation coincident with changes to its replication timing. Such links between chromatin organization and oncogene function underscore the need for mechanistic understanding of how chromatin organization is regulated. The long-term goal is to understand how cells organize chromatin to regulate genomic function to identify and correct the loss of proper organization in cancer. The immediate objective of this application is to define the role and nature of cis elements in the regulation of chromatin organization at the PTN locus. The central hypothesis is that the chromosomal domain in which PTN resides contains DNA elements that allow the domain to be recognized and regulated as an individual functional unit that corresponds to the region that changes replication timing. Preliminary data produced by the applicant and others lead to the formulation of this hypothesis. The rationale that underlies the proposed research is that understanding the role and nature of cis regulation of the PTN chromosomal domain will allow for hypothesis-driven approaches to identify the trans factors responsible for organizing chromatin in the nucleus and will provide insight as to how oncogenes are activated during oncogenesis. The central hypothesis will be tested by pursuing two specific aims: 1) Isolate minimal units of replication timing regulation at the PTN locus; and 2) Determine the role of replication timing in the functional regulation of PTN. To achieve these aims, single-copy segments of the PTN domain will be integrated into a different chromosomal domain with opposite replication timing. Segments that maintain replication-timing regulation will be reduced to minimal units of regulation for the first aim. Under the second aim, PTN promoter activity in segments that do not maintain replication-timing regulation will be compared to activit at the endogenous locus. The expected contributions of the proposed research are to reveal the nature of cis elements regulating chromatin organization and determine their role in the regulation of gene function. These contributions will be significant because identifying regulatory elements is our first step toward uncovering the mechanisms governing cell-type specific chromatin organization and its links to cancer. PUBLIC HEALTH RELEVANCE: The proposed research is relevant to public health because advances in our understanding of the mechanism whereby cells differentially regulate DNA function provide insight as to how that regulation is undermined in cancer. This project is relevant to the mission of National Cancer Institute because understanding the regulation of oncogene function is critical to understanding how oncogenes are activated to cause cancer and what measures should be taken to prevent or reverse their activation in patients.

描述(由申请人提供)：DNA复制、时间和染色质组织的其他协同调节特性在癌症的致癌基因位点上被破坏。Pleiotroin(PTN)是一种在所有恶性胶质瘤中表达的癌基因，在神经分化过程中受转录调控，与其复制时间的变化相一致。染色质组织和癌基因功能之间的这种联系强调了对染色质组织是如何被调节的机械性理解的必要性。长期目标是了解细胞如何组织染色质来调节基因组功能，以识别和纠正癌症中适当组织的丢失。此应用程序的直接目标是定义 顺式元件在调节PTN基因座染色质组织中的作用和性质。中心假设是PTN所在的染色体结构域包含DNA元件，这些DNA元件允许将该结构域识别和调节为与改变复制时间的区域相对应的单个功能单位。申请人和其他人提供的初步数据导致了这一假设的提出。这项研究的理论基础是，了解PTN染色体结构域顺式调控的作用和性质将允许采用假说驱动的方法来确定负责在细胞核中组织染色质的反式因子，并将有助于深入了解癌基因在肿瘤发生过程中是如何被激活的。中心假设将通过追求两个特定的目标来检验：1)分离PTN基因座上复制定时调节的最小单位；2)确定复制定时在PTN功能调节中的作用。为了达到这些目的，PTN结构域的单拷贝片段将以相反的复制时间整合到不同的染色体结构域中。为了第一个目标，维持复制定时调节的片段将减少到最小调节单位。在第二个目标下，不维持复制定时调节的片段中的PTN启动子活性将与内源位置的激活相比较。这项研究的预期贡献是揭示调节染色质组织的顺式元件的性质，并确定它们在基因功能调节中的作用。这些贡献将是重大的，因为确定监管 元素是我们朝着揭示控制细胞类型特定染色质组织及其与癌症的联系的机制迈出的第一步。 公共卫生相关性：拟议的研究与公共健康相关，因为我们对细胞差异调控DNA功能的机制的理解取得进展，为癌症中这种调控是如何破坏的提供了洞察力。这个项目与国家癌症研究所的使命相关，因为了解癌基因功能的调节对于了解癌基因是如何激活导致癌症以及应该采取哪些措施来防止或逆转患者的激活至关重要。