Mechanims of Archaeal Transcription Termination

古菌转录终止机制

基本信息

批准号：
8990012
负责人：
Thomas James Santangelo
金额：
$ 27.13万
依托单位：
COLORADO STATE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2012
资助国家：
美国
起止时间：
2012-02-01 至 2017-12-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8990012
关键词：
Archaea Archaeal RNA Bacteria Biological Assay Cell Differentiation process Complement Complex Conserved Sequence DNA DNA Polymerase I DNA-Directed RNA Polymerase Defect Development Elongation Factor Event Gene Deletion Gene Expression Gene Expression Regulation Gene Proteins Gene-Modified Genes Genetic Genetic Transcription Goals Growth and Development function Homologous Gene Human In Vitro Intrinsic factor Knowledge Malignant Neoplasms Mechanics Modeling Molecular Molecular Biology Monitor Mutagenesis Palpable Phenotype Process Prokaryotic Cells Proteins RNA RNA chemical synthesis Regulation Research Resistance Resolution Role Scheme Signal Transduction Small RNA Stimulus Structure System Techniques Transcription Elongation Transcriptional Regulation Variant Work antitermination attenuation base combat field study flexibility in vivo insight novel premature promoter rRNA Genes research study response termination factor transcription factor transcription termination transcription termination factor I

项目摘要

DESCRIPTION (provided by applicant): Timely and accurate regulation of gene expression is required for proper growth, development, and response to environmental stimuli. A complete understanding of the mechanisms employed to regulate gene expression is necessary to combat the aberrant regulation that underlies many human developmental conditions and ailments, including cancer. The long-term goals of this proposal are to determine the mechanics of RNA synthesis by DNA-dependent multi-subunit RNA polymerases (RNAPs), the regulation imposed on RNAP by conserved protein factors and select template sequences, and to specifically characterize the available mechanisms to halt RNA synthesis and terminate transcription. The proposed experiments take advantage of both an in vivo and in vitro established archaeal transcription system. Archaea offer the advantages of far less complexity but homology in many features of human molecular biology, specifically conservation of RNAP structure and function. We will select, generate, purify and characterize variant RNAPs with modified transcription termination phenotypes. We will further characterize factor-dependent termination, polarity, and the role of such regulation in Archaea. The results so obtained will determine the structures and sequences that support transcription elongation complex stability, describe the mechanics of termination and allow comparisons of termination mechanisms in each Domain, and open a new field of study, archaeal factor-dependent termination.

描述（由申请人提供）：适当的生长，发育和对环境刺激的反应需要及时，准确的基因表达调节。对调节基因表达的机制的完整理解对于打击基于许多人类发育条件和疾病（包括癌症）的异常调节是必要的。该提案的长期目标是确定通过DNA依赖性多亚基RNA聚合酶（RNAP）的RNA合成的机制，该调节是通过保守的蛋白质因子和选择的模板序列对RNAP施加的调节，并专门表征了可用的机制，以阻止RNA合成和终止转录。提出的实验利用了体内和体外建立的古细菌转录系统。古细菌提供了复杂性的优势，但在人类分子生物学的许多特征中，特别是对RNAP结构和功能的保存。我们将选择，生成，纯化和表征具有修改的转录终止表型的变体RNAP。我们将进一步表征因子依赖性终止，极性和这种调节在古细菌中的作用。如此获得的结果将确定支持转录伸长复合稳定性的结构和序列，描述终止的机制并允许每个域中的终止机制进行比较，并打开一个新的研究领域，即细分因子依赖性终止。