STRUCTURAL HOMOLOGY BETWEEN RAP30 DMA BINDING DOMAIN & LINKER HISTONE H5

通过多核NMR解析Rap 30-DBD的结构， 揭示了一些组成部分之间的结构关系， 原核和真核RNA聚合酶复合体。 （Caroline M. Groft，Sacha N. Uljon，Rong Wang，and米尔顿H. 03 The Dog（1998） 95，10872-10877）。 原核生物和真核生物之间的类比 在大肠杆菌sigma 70和Rap 30之间构建嵌合体 为了克服前者的溶解性问题， 排除了结构分析。 这一目标现已实现， 已经生产出高度可溶形式的σ 70， 正在筛选活动。 sigma 70第4区的置换 Rap 30-DBD导致可溶形式的sigma 70表明 σ 70溶解度的决定因素可能存在于 包含区域4的60个氨基酸。 诱变/质谱 正在进行分析以制备σ 70的突变体， 固有可溶并保持天然活性。

The structure of Rap30-DBD was solved by multi-nuclear NMR, revealing some of the structural relationship between components of the prokaryotic and eukaryotic RNA polymerase complex. (Caroline M. Groft, Sacha N. Uljon, Rong Wang, and Milton H. Werner (1998) PNAS USA 95, 10872-10877). The analogy between prokaryotes and eukaryotes has been extended to construct a chimera between E coli sigma70 and Rap30 in order to overcome solubility problems in the former which have precluded structural analysis. This has now been accomplished and highly soluble form of sigma70 has been produced and is presently being screened for activity. The replacement of region 4 of sigma70 with the Rap30-DBD that resulted in a soluble form of sigma70 suggests that the determinants of solubility of sigma70 are likely to reside in the 60 amino acids comprising region 4. Mutagenesis/mass spec analysis is under way to prepare mutants of sigma70 that are inherently soluble and retain native activity.