ROGER KORNBERG PRT TIME

该子项目是利用 由NIH/NCRR资助的中心赠款提供的资源。子项目和 研究者（PI）可能从另一个NIH来源获得主要资金， 因此可以在其他CRISP条目中表示。列出的机构是 中心，不一定是研究者的机构。 拟议研究的目标是确定RNA聚合酶II及其与核酸和辅助蛋白质因子的复合物在原子分辨率下的X射线结构。该问题是具有挑战性的，因为单独的聚合酶包含15种多肽，总质量接近600，000道尔顿，并且添加辅助因子使多肽的数量和蛋白质质量加倍。解决这个问题需要结合生物化学研究，电子和X射线晶体学。现在有八种类型的RNA聚合酶II晶体，天然酶的晶体，与DNA的共晶体，与RNA的共晶体，与转录活性状态的DNA和RNA的共晶体，与TFIIB的共晶体，与TFIIE的共晶体，与额外的聚合酶亚基RPB 4和RPB 7的共晶体，以及与紧密结合抑制剂α-鹅膏蕈碱的共晶体。最近的进展包括低分辨率信封和相位到6.5 º，以及最近收集的几个新的重原子簇与RNA Pol II复合。现在的预测是非常好的快速实现高分辨率的结构。

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.