Molecular Characterization of Transcriptional Activation and Repression in Respon

描述(由申请人提供)：适当的转录调控是确保细胞对细胞内提示和环境变化做出适当反应的关键。基因表达的时间或空间缺陷直接影响细胞的生长和分裂、对压力和能源的反应以及相邻细胞之间的交流。在高等真核生物中，转录在控制分化为特定类型的细胞方面起着至关重要的作用，当错误引导时，这一过程可能导致细胞生长失控和肿瘤形成。了解转录是如何调控的一个关键部分是对转录周期中各个步骤中发生的一整套因素和事件有充分的了解。虽然已经确定了许多关键因素，但我们缺乏在分子方面获得对实现目标基因激活或抑制所需步骤的完整看法的联系。我们理解上的一个主要差距是由于缺乏一种基于感兴趣的基因组区域而不是调控蛋白本身的分离技术。通过采用金斯顿实验室最近开发的方法来分离基因组的特定区域及其相关蛋白质，我们将使用果蝇黑腹果蝇详细研究特定基因如何在热休克形式的环境胁迫下被激活和抑制。果蝇强烈调节的热休克反应非常适合于分析转录的受控、快速激活和抑制，并揭示人类细胞中也存在的转录调节的一般机制。这项研究将允许无偏见地识别新的转录因子，并允许根据它们的时间角色来表征已知的因子。热休克途径的阐明将提供有价值的新信息，将允许重组现有的转录调控模式，并将为未来的功能研究打开大门。通过对应激反应基因正常激活和抑制的分子细节的描述，我们可以开始了解这些过程是如何在癌症中被破坏的。)

DESCRIPTION (provided by applicant): The proper regulation of transcription is essential to ensure that cells respond appropriately to intracellular cues and environmental variations. Temporal or spatial defects in gene expression directly impact cell growth and division, responses to stress and energy sources, and communication between neighboring cells. In higher eukaryotes, transcription plays a vital role in controlling differentiation into specific cell types, a process that can lead to uncontrolled cell growth and tumor formation when misdirected. A crucial part of understanding how transcription is regulated is to have full knowledge of the complete set of factors and events that take place throughout the various steps in the transcription cycle. While many key factors have been identified, we lack the links to gain a complete view in molecular terms of the steps required to achieve activation or repression of a target gene. A major gap in our understanding has been due to the absence of an isolation technique based on the genomic region of interest rather than the regulatory proteins themselves. By adapting a method recently developed by the Kingston laboratory to isolate specific regions of the genome along with their associated proteins, we will study in detail how specific genes are activated and repressed in response to environmental stress in the form of heat shock using the fruit fly Drosophila melanogaster. The robustly regulated heat shock response in Drosophila is ideally suited to analyze the controlled, rapid activation and repression of transcription and to uncover general mechanisms of transcriptional regulation that also occur in human cells. This study will allow for unbiased identification of new transcription factors, and permit the characterization of known factors according to their temporal roles. The elucidation of the heat shock pathway will provide valuable new information that will allow for the restructuring of current models of transcriptional regulation and will open the door for future functional studies. By characterizing in molecular detail the normal activation and repression of stress response genes, we can begin to understand how these processes are disrupted in cancers. )