STRUCTURE OF AN ARCHAEAL TRANSCRIPTION COMPLEX

古菌转录复合体的结构

• 批准号: 6179908
• 负责人: MICHAEL S BARTLETT
• 金额: $ 3.75万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN DIEGO
• 依托单位国家: 美国
• 财政年份: 1999
• 资助国家: 美国
• 起止时间: 1999-06-01 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/6179908
• 关键词: Archaea; DNA binding protein; DNA directed RNA polymerase; RNA binding protein; crosslink; genetic promoter element; genetic transcription; microorganism genetics; protein structure; transcription factor

The archaea, or archaebacteria, were recognized as a form of life distinct from bacteria and eucarya just two decades ago. Studies of the archaeal transcription apparatus indicate that the archaeal system is highly homologous to the system in eucarya. However, little is known about the detailed mechanism of transcription or its regulation in the archaea. The experiments in this project attempt to address these questions by defining the topographical structure of the transcription complex from the hyperthermophilic archaeon Pyrococcus furiosus. The arrangement of the RNA polymerase subunits relative to promoter DNA and to the RNA transcript will be determined at defined steps in transcription using DNA-protein and RNA-protein photochemical cross- linking methods. In addition, the interactions between RNAP and the transcription initiation factors TBP and TFB will be investigated by protein-protein footprinting and protein cross-linking methods. Finally, the function of a putative novel transcription factor, homologous to TFB, will be investigated through binding and transcription assays with a variety of P. furiosus promoters. These experiments should provide an excellent overview of the structure of the archaeal transcription apparatus, and will provide a foundation for studying the regulation of transcription in this biological domain. Because of the current scarcity of basic knowledge concerning archaeal biology, the experiments in this proposal are very likely to result in discovery and to yield novel observations. In addition, the homology between archaeal and eucaryal transcription implies that conclusions from these experiments will be directly applicable to the study of eucaryal transcription. Therefore, this work may provide health benefits arising from applications to human inherited or acquired disease that originate from aberrant gene transcription. With a better understanding of eucaryal transcriptional mechanism, points of leverage for disease control may be identified and exploited.

古生菌或古细菌被认为是一种生命形式。 与二十年前的细菌和真核生物截然不同。对中国传统文化的研究 古生菌转录装置表明古生菌系统是 与尤卡里亚的系统高度同源。然而，人们对此知之甚少 关于转录的详细机制或其在 古生菌。本项目中的实验试图解决这些问题 通过定义转录的地形结构来提问 来自嗜热古生菌的复合体。这个 RNA聚合酶亚基相对于启动子DNA和 中定义的步骤确定到RNA转录本 利用DNA-蛋白质和RNA-蛋白质光化学交叉转录 链接方法。此外，RNAP与 转录起始因子TBP和TFB将通过 蛋白质-蛋白质足迹和蛋白质交联法。 最后，推测一种新的转录因子的功能， 与TFB同源的基因将通过结合和 用不同的P.Furiosus启动子进行转录检测。这些 实验应该提供一个很好的结构概述 古生菌转录装置，并将为 研究这个生物结构域中转录的调节。 因为目前关于古生物的基本知识匮乏 生物学，这项提议中的实验很可能导致 发现并产生新的观察结果。此外，同源性 古文字和真彩色文字之间的差异意味着结论 这些实验的结果将直接应用于研究 欧几里亚语转录。因此，这项工作可能会提供健康 应用于人类遗传或后天获得的好处 由基因转录异常引起的疾病。拥有更好的 了解欧几里亚尔转录机制、杠杆点 可以识别和开发用于疾病控制的。