LOCUS CONTROL REGION HSI,II AND HGH EXPRESSION

位点控制区 HSI、II 和 HGH 表达

• 批准号: 6176344
• 负责人: BRIAN M SHEWCHUK
• 金额: $ 3.92万
• 依托单位: UNIVERSITY OF PENNSYLVANIA
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-09-01 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/6176344
• 关键词: binding proteins; gel mobility shift assay; gene expression; genetic mapping; genetic promoter element; genetic regulation; genetic regulatory element; genetically modified animals; laboratory mouse; molecular cloning; somatotropin

The pituitary- and placenta-specific genes of the human growth hormone (hGH) gene cluster are vital to normal growth and physiology during development and in the adult. Misregulation of the hGh gene leads to significant pathological consequences that illustrate its importance in postnatal development. Furthermore, the organization of genes in the cluster and its association with a dominant distal regulatory element, the locus control region (LCR), make it a valuable model for the study of genes in higher eukaryotes that are organized into discrete genomic regulatory domains. The work proposed here will address important questions about the modulation of gene expression during in utero development with a focus on the mechanisms by which LCRs are involved as major determinants in this process. The aims of the proposed research are to study the organization and identity of a subset of regulatory determinants in the hGH LCR ( HSI, II) and their role in the establishment ant maintenance of an hGH transgene in a transcriptionally active state during pituitary development. The region of the LCR involved in the transcriptional activation of the hGH gene in the pituitary has been identified by its DnaseI hypersensitivity (HSI,II) and is the focus of this proposal. In aim I, using a transgenic mouse system, the precise locations of LCR activity will be mapped in the HSI, II region. In Aim II, the discrete elements involved in HSI,II region. In Aim II, the discrete elements involved in HSI,II activity and their corresponding protein factors will be identified and characterized using in vitro protein binding and foot printing assays. There is a strong potential for the discovery of novel regulatory proteins in this Aim, as there are no recognition sequences for known factors in the HSI,II region. Finally, Aim III will determine whether HSI,II is required the maintenance as well as the initial activation of hGH gene expression during pituitary somatotrope cell lineage progression in a transgenic mouse system using a developmentally-regulated conditional deletion of HSI,II. The outcome of this research, if successful , will result in novel insight into the developmental control of human growth hormone gene cluster, the mechanism by which LCRs function, and how gene expression profiles are established and maintained in specific cell lineages during development.

人类生长发育的垂体和胎盘特异性基因 激素（hGH）基因簇对于正常生长和生理是至关重要 在发育期和成年期。 hGh基因的失调 导致了严重的病理后果， 对产后发育的重要性。 此外，该组织 基因簇中的基因及其与显性远端 调控元件，基因座控制区（LCR），使其成为一个 研究高等真核生物基因的有价值的模型， 组织成离散的基因组调控域。 工作 这里提出的将解决有关调制的重要问题 在子宫内发育过程中的基因表达，重点是 LCR作为主要决定因素参与其中的机制 过程 拟议研究的目的是研究 组织和身份的一个子集的监管决定因素， hGH LCR（HSI，II）及其在建立蚂蚁中的作用 维持hGH转基因处于转录活性状态 在垂体发育过程中。 LCR的区域参与了 垂体中hGH基因的转录激活已经被证实是 通过其DnaseI超敏反应（HSI，II）鉴定， 这一提议。 在目的I中，使用转基因小鼠系统， 在第二阶段的《人类健康指数》中，将绘制LCR活动的精确位置图 地区 在Aim II中，离散元涉及HSI，II区域。 在目标II中，涉及HSI、II活动和 它们相应的蛋白质因子将被鉴定， 使用体外蛋白质结合和足迹法 测定。 有很大的潜力去发现新的东西 调节蛋白在这个目标，因为没有识别 HSI，II区域中已知因子的序列。 第三，目标 将决定是否需要HSI，II维护以及 垂体生长激素基因表达的初始激活 转基因小鼠系统中生长激素细胞谱系进展 使用发育调节的HSI的条件性缺失，II. 这项研究的结果，如果成功，将导致新的 人类生长激素基因发育调控研究进展 集群，LCR的功能机制，以及基因如何 在特定细胞中建立并维持表达谱 发展过程中的血统。