Transcription: Mechanism and Regulation

 描述（由申请人提供）：拟议的研究是我们长期努力的一部分，使用细菌模型系统研究转录及其调控的基本机制。基因调控的适当控制对于生物体发育、细胞对环境信号的反应以及疾病状态的预防至关重要。基因表达的第一步，转录，是由多亚基RNA聚合酶（RNAP）进行的，这些酶在序列、结构和功能上都是保守的。因此，对细菌中转录的详细机制理解有助于理解所有生物体中的转录。拟议的研究将采用综合方法，结合遗传学，生物化学，结构生物学，基因组学，和基于高通量测序的方法来（1）理解将细胞状态的变化与RNAP活性的变化联系起来的调节机制的多样性和（2）获得转录的定量理解，定义转录每一步的机制基础，并赋予能力，从基因的DNA序列内容预测转录输出。我们通过开发新的方法来理解DNA序列和RNAP功能之间的界面，并重新审视长期以来对RNAP功能的概念，以解决这些领域的问题。

 DESCRIPTION (provided by applicant): The proposed studies are part of our long-range effort to study the fundamental mechanisms of transcription and its regulation using bacterial model systems. Proper control of gene regulation is critical for organismal development, cellular response to environmental signals, and the prevention of disease states. The first step in gene expression, transcription, is carried out by multi-subunit RNA polymerases (RNAPs) that are conserved in sequence, structure and function from bacteria to humans. Thus, a detailed mechanistic understanding of transcription in bacteria facilitates an understanding of transcription in all organisms. The proposed studies will apply a comprehensive approach combining genetics, biochemistry, structural biology, genomic, and high- throughput sequencing based methods to (1) understand the diversity of regulatory mechanisms that link changes to cellular state to changes in the activity of RNAP and (2) obtain a quantitative understanding of transcription that defines the mechanistic basis for each step of transcription and confers the ability to predict transcriptional output from the DNA sequence content of a gene. We tackle these areas by developing new methodologies to understand the interface between DNA sequence and RNAP function, and to reexamine long-held notions of RNAP function for unexpected paradigms or unknown biology.