Mechanims of Archaeal Transcription Termination

古菌转录终止机制

• 批准号: 8221794
• 负责人: Thomas James Santangelo
• 金额: $ 27.83万
• 依托单位: OHIO STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-02-01 至 2016-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8221794
• 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; Genes; Genetic; Genetic Transcription; Goals; Growth and Development function; Homologous Gene; Human; In Vitro; Intrinsic factor; Knowledge; Malignant Neoplasms; Mechanics; Modeling; Modification; Molecular; Molecular Biology; Monitor; Mutagenesis; Palpable; Phenotype; Process; Prokaryotic Cells; Proteins; RNA; RNA chemical synthesis; Regulation; Relative (related person); 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

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. PUBLIC HEALTH RELEVANCE: 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 results so obtained will determine the structures and sequences that support transcription elongation complex stability, describe the mechanisms 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。为了对抗许多人类发育状况和疾病（包括癌症）的异常调节，必须全面了解用于调节基因表达的机制。该提案的长期目标是确定 DNA 依赖性多亚基 RNA 聚合酶 (RNAP) 合成 RNA 的机制、保守蛋白因子和选择模板序列对 RNAP 的调节，并具体表征停止 RNA 合成和终止转录的可用机制。所提出的实验利用了体内和体外建立的古菌转录系统。古细菌的优点是复杂性低得多，但在人类分子生物学的许多特征上具有同源性，特别是 RNAP 结构和功能的保守性。我们将选择、生成、纯化和表征具有修饰转录终止表型的变异 RNAP。我们将进一步描述古细菌中因子依赖性终止、极性以及此类调节的作用。如此获得的结果将确定支持转录延伸复合物稳定性的结构和序列，描述终止机制并允许比较每个域中的终止机制，并开辟一个新的研究领域，即古菌因子依赖性终止。 公共卫生相关性：为了对抗许多人类发育状况和疾病（包括癌症）的异常调节，必须全面了解用于调节基因表达的机制。该提案的长期目标是确定 DNA 依赖性多亚基 RNA 聚合酶 (RNAP) 合成 RNA 的机制、保守蛋白因子和选择模板序列对 RNAP 的调节，并具体表征停止 RNA 合成和终止转录的可用机制。如此获得的结果将确定支持转录延伸复合体稳定性的结构和序列，描述终止机制并允许比较每个域中的终止机制，并开辟一个新的研究领域，即古菌因子依赖性终止。