Transcription: Mechanism and Regulation

 描述(由申请人提供)：拟议的研究是我们利用细菌模型系统研究转录及其调控的基本机制的长期努力的一部分。对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，即转录，是由多亚单位RNA聚合酶(RNAP)进行的，RNAP在序列、结构和功能上从细菌到人类都是保守的。因此，对细菌转录的详细机制了解有助于理解所有生物体的转录。拟议的研究将综合应用遗传学、生物化学、结构生物学、基因组学和高通量测序方法，以(1)了解将细胞状态的变化与RNAP活性变化联系起来的调控机制的多样性，以及(2)对转录的定量理解，定义转录的每一步的机制基础，并赋予根据基因的DNA序列内容预测转录输出的能力。我们通过开发新的方法来解决这些领域，以了解DNA序列和RNAP功能之间的接口，并重新检查长期持有的RNAP功能的概念，以寻找意外的范例或未知的生物学。

 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.