Sousa R, Chung Y J, Rose J P, Wang B C
Department of Biological Sciences, University of Pittsburgh, Pennsylvania 15260.
Nature. 1993 Aug 12;364(6438):593-9. doi: 10.1038/364593a0.
The crystal structure of T7 RNA polymerase reveals a molecule organized around a cleft that can accommodate a double-stranded DNA template. A portion (approximately 45%) of the molecule displays extensive structural homology to the polymerase domain of Klenow fragment and more limited homology to the human immunodeficiency virus HIV-1 reverse transcriptase. A comparison of the structures and sequences of these polymerases identifies structural elements that may be responsible for discriminating between ribonucleotide and deoxyribonucleotide substrates, and RNA and DNA templates. The relative locations of the catalytic site and a specific promoter recognition residue allow the orientation of the polymerase on the template to be defined.
T7 RNA聚合酶的晶体结构显示,该分子围绕着一个可容纳双链DNA模板的裂隙排列。该分子的一部分(约45%)与Klenow片段的聚合酶结构域具有广泛的结构同源性,与人类免疫缺陷病毒HIV-1逆转录酶的同源性则较为有限。对这些聚合酶的结构和序列进行比较,可确定可能负责区分核糖核苷酸和脱氧核糖核苷酸底物以及RNA和DNA模板的结构元件。催化位点和特定启动子识别残基的相对位置使得能够确定聚合酶在模板上的方向。