MACCHESS PROGRAM FOR LARGE UNIT CELLS: HIV

针对大型单元格的 MACCHESS 程序：HIV

• 批准号: 7357714
• 负责人: JIANHUA FU
• 金额: $ 8.29万
• 依托单位: CORNELL UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-07-01 至 2007-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7357714
• 关键词: MACCHESS; PROGRAM; LARGE; UNIT; CELLS

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. This lab has been working towards a solution of the cocrystal structure of RNA polymerase II-TFIIF complex. The transcription system of RNA polymerase II (Pol II) has emerged as an enormous protein apparatus consisting of the polymerase at its heart surrounded by layers of controlling factors. With recent crystallographic advances, it is now feasible to structurally characterize the apparatus in various complex forms, so we can understand Pol II-factor(s) interactions that modulate the activities of Pol II. Indeed, having a "big picture" of how proteins interact in the Pol II machinery (a total of 50 polypeptides) and knowing the "language" of their communication are the next challenges facing the structural biology of transcription. Using MacCHESS F1 and F2 stations, we have collected two native data sets to 7.5 ¿ from Pol II-TFIIF cocrystal with good statistics. Since the Pol II in the complex has been determined, we are now analyzing the data using protein phases generated from the polymerase model. Densities additional to Pol II model are clearly visible in three locations that could be arising from the TFIIF subunit, Tfg1, Tfg2 and Tfg3. More recently, we collected low-resolution data from cocrystals of another Pol II complex: the Pol II-capping enzyme complex. The capping enzyme catalyzes the formation of a special structure (the 5' cap) at the 5'-end of nascent RNA transcripts made by Pol II and is essential for cell vitality. Using combined phases from the Pol II model and an SAS set, we observed densities additional to the model structure and confined to a region adjacent to a helical domain of Pol II largest subunit Rpb1.

该子项目是利用NIH/NCRR资助的中心赠款提供的资源的许多研究子项目之一。子弹和调查员（PI）可能已经从其他NIH来源获得了主要资金，因此可以在其他清晰的条目中代表。列出的机构是针对该中心的，这不是调查人员的机构。该实验室一直致力于RNA聚合酶II-TFIIF复合物的共晶结构的溶液。 RNA聚合酶II（POL II）的转录系统已成为一种巨大的蛋白质设备，该设备由聚合酶在其心脏的聚合酶组成，周围是由控制因子层的层。随着最近的晶体学进步，现在可以在结构上以各种复杂形式来表征该设备是可行的，因此我们可以理解调节Pol II活性的Pol II因子相互作用。的确，拥有蛋白质如何在Pol II机械（总共50种多肽）中相互作用并了解其交流的“语言”是转录结构生物学面临的下一个挑战。使用MacChess F1和F2站，我们从具有良好统计数据的Pol II-Tfiif Cocrystal中收集了两个天然数据集。由于已经确定了复合物中的POL II，因此现在使用从聚合酶模型产生的蛋白质相分析数据。在TFIIF亚基，TFG1，TFG2和TFG3可能引起的三个位置中，可以清楚地看到Pol II模型的密度。最近，我们从另一个Pol II复合物的共晶（Pol II限制酶复合物）中收集了低分辨率数据。上限酶催化了POL II的新生RNA转录本的5'末端的特殊结构（5'CAP）的形成，对于细胞活力是必不可少的。使用POL II模型和SAS集的组合相，我们观察到模型结构以外的密度，并局限于Pol II最大亚基RPB1的螺旋结构域附近的区域。