ROGER KORNBERG PRT TIME

这个子项目是利用由NIH/NCRR资助的中心拨款提供的资源的许多研究子项目之一。子项目和调查员(PI)可能从另一个NIH来源获得了主要资金，因此可能会出现在其他CRISE条目中。列出的机构是针对中心的，而不一定是针对调查员的机构。这项拟议研究的目标是在原子分辨率下确定RNA聚合酶II及其与核酸和辅助蛋白质因子的络合物的X射线结构。这个问题很有挑战性，因为仅聚合酶就由15个多肽组成，总质量近60万道尔顿，加上辅助因子，多肽的数量和蛋白质质量都翻了一番。这个问题的解决需要生化研究、电子和X射线结晶学的结合。目前已有八种类型的RNA聚合酶II晶体，它们是天然酶的晶体，以及与DNA、与RNA、与DNA和RNA处于转录活性状态、与TFIIB、与TFIIE、与附加聚合酶亚基RPB4和RPB7以及与紧密结合的抑制剂a-Amanitin的共晶体。最近的进展包括低分辨率包络和相位到6.5°，以及最近收集的几个新的与RNA POL II络合的重原子团簇。现在预测非常好的是快速实现高分辨率结构。

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. The goal of the proposed research is to determine the X-ray structures of RNA polymerase II and of its complexes with nucleic acids and auxiliary protein factors at atomic resolution. The problem is challenging, since the polymerase alone comprises 15 polypeptides with a total mass of nearly 600,000 Dalton, and addition of the auxiliary factors doubles both the number of polypeptides and the protein mass. Solution of the problem requires a combination of biochemical studies, electron and X-ray crystallography. Eight types of RNA polymerase II crystal are now in hand, crystals of the native enzyme, and cocrystals with DNA, with RNA, with both DNA and RNA in the transcriptionally active state, with TFIIB, with TFIIE, with the additional polymerase subunits RPB4 and RPB7, and with the tight-binding inhibitor a-amanitin. Recent progress includes a low resolution envelope and phases to 6.5 ¿¿ and the recent collection of several novel heavy atom cluster complexed with RNA Pol II. The prognosis is now very good for the rapid achievement of a high resolution structure.