STRUCTURE DETERMINATION OF RNA POLYMERASE II BY XRAY DIFFRACTION METHOD

该项目的目标是确定X光结构 RNA聚合酶II及其与核酸和核酸的络合物 原子分辨率的辅助蛋白质因子。问题是 具有挑战性，因为仅聚合酶就由15个多肽组成 总质量近60万道尔顿，加上辅助剂 因子使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结合生化研究， 电子和X射线结晶学。在过去的十年里，我们已经 完成了酵母RNA聚合酶II的生化解剖 转录系统，并测定了RNA聚合酶II 16岁时的结构二维(2D)电子的分辨率 显微结晶学。最近，我们已经获得了三维晶体 这一数字已衍变至3.1并且此后已将相分解到6.5 采用MIRAS方法。本项目的成果将为今后的研究提供依据 为了理解大量的生化和获取数据 在过去的30年里，转录在不同的系统中积累起来。

The goal of this project 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 Daltons, 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. During the past decade, we have completed the biochemical dissection of the yeast RNA polymerase II transcription system, and we have determined RNA polymerase II structure at 16
resolution by two-dimensional (2D) electron microscopy crystallography. Recently, we have obtained 3-D crystals that diffracted to 3.1
and have thereafter resolved phases to 6.5
using MIRAS method. The result of this project will provide a basis for understanding a vast body of biochemical and getetic data on transcription accumulated in diverse systems over the past 30 years.