Structural basis of DNA recognition by PCG2 reveals a novel DNA binding mode for winged helix-turn-helix domains.
Structural basis of DNA recognition by PCG2 reveals a novel DNA binding mode for winged helix-turn-helix domains.
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PCG2 识别 DNA 的结构基础揭示了翼状螺旋-转角-螺旋结构域的新型 DNA 结合模式。
DOI：
10.1093/nar/gku1351
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发表时间：
2015-01
影响因子：
14.9
通讯作者：
Peng YL
中科院分区：
文献类型：
--
作者：
Liu J;Huang J;Zhao Y;Liu H;Wang D;Yang J;Zhao W;Taylor IA;Peng YL
MBP 1家族蛋白是MBF细胞周期转录因子复合物的DNA结合亚基，并且含有N末端翼螺旋-转角-螺旋（wHTH）DNA结合结构域（DBD）。虽然酿酒酵母MBP 1的DNA结合机制已被广泛研究，但其他MBP 1家族蛋白的结构框架和DNA结合模式仍有待揭示。在这里，我们确定了与MCB-DNA结合的PCG 2（MBP 1的Magnaporthe β直系同源物）的DBD的晶体结构。结构分析表明，PCG 2-DBD中的翼区、20-环、螺旋A和螺旋B是DNA结合的重要元件。与先前表征的wHTH蛋白不同，PCG 2-DBD利用翼和螺旋B结合MCB-DNA的小沟和大沟，而20环和螺旋A与DNA非特异性相互作用。值得注意的是，发现翼内的两个谷氨酰胺Q89和Q82通过形成氢键相互作用识别MCB核心CGCG序列。进一步的体外测定证实了Q89和Q82在DNA结合中的重要作用。这些数据共同表明，MBP 1同源物PCG 2采用了一种不寻常的模式结合靶DNA，并证明了wHTH结构域的多功能性。
The MBP1 family proteins are the DNA binding subunits of MBF cell-cycle transcription factor complexes and contain an N terminal winged helix-turn-helix (wHTH) DNA binding domain (DBD). Although the DNA binding mechanism of MBP1 from Saccharomyces cerevisiae has been extensively studied, the structural framework and the DNA binding mode of other MBP1 family proteins remains to be disclosed. Here, we determined the crystal structure of the DBD of PCG2, the Magnaporthe oryzae orthologue of MBP1, bound to MCB–DNA. The structure revealed that the wing, the 20-loop, helix A and helix B in PCG2–DBD are important elements for DNA binding. Unlike previously characterized wHTH proteins, PCG2–DBD utilizes the wing and helix-B to bind the minor groove and the major groove of the MCB–DNA whilst the 20-loop and helix A interact non-specifically with DNA. Notably, two glutamines Q89 and Q82 within the wing were found to recognize the MCB core CGCG sequence through making hydrogen bond interactions. Further in vitro assays confirmed essential roles of Q89 and Q82 in the DNA binding. These data together indicate that the MBP1 homologue PCG2 employs an unusual mode of binding to target DNA and demonstrate the versatility of wHTH domains.
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影响因子：
11.4
作者：
MIYAMOTO, M;TANAKA, K;OKAYAMA, H
通讯作者：
OKAYAMA, H
影响因子：
3.2
作者：
Chernatynskaya, Anna V.;Deleeuw, Lynn;Lane, Andrew N.
通讯作者：
Lane, Andrew N.
DOI：
10.1107/s0907444909052925
发表时间：
2010-02
期刊：
Acta crystallographica. Section D, Biological crystallography
影响因子：
--
作者：
Adams PD;Afonine PV;Bunkóczi G;Chen VB;Davis IW;Echols N;Headd JJ;Hung LW;Kapral GJ;Grosse-Kunstleve RW;McCoy AJ;Moriarty NW;Oeffner R;Read RJ;Richardson DC;Richardson JS;Terwilliger TC;Zwart PH
通讯作者：
Zwart PH
影响因子：
64.8
作者：
CLARK, KL;HALAY, ED;BURLEY, SK
通讯作者：
BURLEY, SK
影响因子：
5.8
作者：
Larkin, M. A.;Blackshields, G.;Higgins, D. G.
通讯作者：
Higgins, D. G.