Posttranscriptional regulation of gene expression in eukaryotes

描述（由申请人提供）：该申请涉及广泛的挑战领域（06）使能技术和特定的挑战主题，06- gm -112：分子和细胞动力学技术。生物学研究的一个中心问题是理解基因组是如何被解释以确定生物形态和功能的巨大多样性的。此外，为了解剖和预防人类疾病的发展，确定在解释遗传信息期间可能发生的错误调控是至关重要的。虽然已经确定基因表达调控的许多重要步骤发生在转录水平上，但越来越清楚的是，转录后加工对于真核细胞中基因表达的精确控制也是必不可少的。例子包括rna介导的沉默通路的发现，该通路控制mRNA的翻译或衰变，并在生物体发育或癌症等疾病的发展过程中发挥关键作用。然而，与我们对转录控制机制的良好理解相比，我们对转录后调控程序知之甚少。这部分是由于缺乏技术，使我们能够以动态和非侵入性的方式分析转录后的控制步骤。在本应用中，我们将解决这一挑战，并提出开发和应用新的标记方法，这将使我们能够在系统范围内以高灵敏度测量单个rna和蛋白质的周转，而不会干扰细胞环境。这将使我们第一次能够将转录中发生的变化与mRNA稳定性、翻译和蛋白质周转的测量结合起来。总之，这将使我们能够对不同生长条件和不同细胞类型下基因表达的调控有一个全面的概述。从长远来看，我的实验室将探索从这些研究中出现的新范式，并将阐明新调控反应的分子机制。这些研究将为决定基因表达模式的细胞动力学的基本原理提供关键的见解，并帮助我们了解人类疾病中发生的缺陷。

DESCRIPTION (provided by applicant): This application addresses broad Challenge Area (06) Enabling Technologies and specific Challenge Topic, 06-GM-112: Molecular and Cellular Dynamics Technologies. A central problem in biological research is to understand how the genome is interpreted to determine the large diversity of organismal form and function. In addition, it is critical to identify misregulations that can occur during the interpretation of the genetic information in order to dissect and prevent the development of human diseases. While it has been well established that many important steps in the regulation of gene expression occur at the level of transcription, it has become increasingly clear that posttranscriptional processing is also essential for the precise control of gene expression in eukaryotic cells. Examples include the discovery of RNA-mediated silencing pathways that control mRNA translation or decay and play critical roles during organismal development or during the development of diseases such as cancer. However, in contrast to our excellent understanding of transcriptional control mechanisms, very little is still known about post-transcriptional regulatory programs. This is in part due to the lack of technologies that would enable us to analyze post-transcriptional control steps in a dynamic and non-invasive fashion. In this application, we will address this challenge and propose the development and application of novel labeling methods that will allow us to measure on a system-wide level the turnover of individual RNAs and proteins with high sensitivity and without perturbation of the cellular environment. For the first time, this will allow us to integrate changes that occur in transcription with measurements of mRNA stability, translation and protein turnover. Together, this will allow us to develop a comprehensive overview of the regulation of gene expression under various growth conditions and in different cell types. In the long run, my laboratory will explore new paradigms that emerge from these studies and will elucidate the molecular mechanism underlying novel regulatory responses. These studies will provide critical insight into the fundamental principles that determine the cellular dynamics of gene expression patterns and help us to understand defects that occur in human diseases. PUBLIC HEALTH RELEVANCE: In all eukaryotes, the tight regulation of gene expression is essential for cells to control proliferation, differentiation or development, and errors in any step of the gene expression program can have severe consequences leading to cell death or disease. A critical step in the control of gene expression is the control of mRNA and protein abundance by the regulation of their decay. Therefore, a better understanding of the regulation of mRNA and protein turnover is essential both for the understanding of fundamental cellular processes and the development of novel therapies for human diseases.