Solution structure of the DNA-binding domain of Cd2-GAL4 from S. cerevisiae
Solution structure of the DNA-binding domain of Cd2-GAL4 from S. cerevisiae
复制标题
酿酒酵母 Cd2-GAL4 DNA 结合域的溶液结构
作者：
J. Baleja;R. Marmorstein;S. Harrison;G. Wagner
GAL 4蛋白激活酿酒酵母1中半乳糖利用所需基因的转录。该蛋白质由881个氨基酸组成，当与半乳糖上游激活序列（UASG）2-5中存在的约两倍对称DNA位点之一结合时，为二聚体。在这里，我们使用二维NMR光谱来确定GAL 4的氨基末端片段（残基1-65）的结构。该片段是溶液中的单体，作为二聚体特异性结合含UASG的DNA。残基9-40形成了一个明确的，紧凑的球状簇，而残基1-8和41-66显示出相当大的构象迁移率在DNA的情况下。紧凑的结构域包含一个基序，其中六个半胱氨酸，位于两个对称相关的螺旋/延长链单位连接一个长环，协调两个中心锌离子，形成一个双金属硫醇盐簇6 -11。在我们的大部分工作中，锌被NMR活性的113 Cd取代，因此结构参数来自Cd 2蛋白。GAL 4 DNA结合结构域的结构代表了一种新的DNA结合基序。使用NMR技术观察到溶液中的紧密结构域的构象与使用晶体学技术观察到与DNA结合的N-末端片段的中心核心的构象基本相同12。因此，DNA结合结构域的核心在结合DNA时几乎没有变化。
THE GAL4 protein activates transcription of the genes required for galactose utilization in Saccharomyces cerevisiae 1. The protein, consisting of 881 amino acids, is dimeric when bound to one of the approximately twofold symmetrical DNA sites present in the galactose upstream activating sequence (UASG)2–5. Here we use two-dimensional NMR spectroscopy to determine the structure of an amino-terminal fragment of GAL4 (residues 1-65). This fragment, a monomer in solution, binds as a dimer specifically to UASG-containing DNA. Residues 9-40 form a well defined, compact globular cluster, whereas residues 1-8 and 41-66 show considerable conformational mobility in the absence of DNA. The compact domain contains a motif in which six cysteines, located on two symmetrically related helix/extended strand units connected by a long loop, coordinate two central zinc ions, forming a bimetal-thiolate cluster6–11. The zincs were replaced by NMR-active113Cd in most of our work and structural parameters are therefore derived from the Cd2-protein. The structure obtained for the GAL4 DNA-binding domain represents a novel DNA-binding motif. Essentially the same conformation is observed for the compact domain in solution using NMR techniques as was seen for the central core of the N-terminal fragment bound to DNA using crystallographic techniques12. Thus, the core of the DNA-binding domain changes little upon binding DNA.
影响因子：
56.9
作者：
KEEGAN, L;GILL, G;PTASHNE, M
通讯作者：
PTASHNE, M
GAL4 转录因子 DNA 结合域的 Zn(II)2 和 113Cd(II)2 衍生物的 1H NMR 共振序列分配揭示了特异性 DNA 识别的新结构基序。
影响因子：
2.9
作者：
Pan,T;Coleman,JE
通讯作者：
Coleman,JE