The solution structure of the N-terminal zinc finger of GATA-1 reveals a specific binding face for the transcriptional co-factor FOG
The solution structure of the N-terminal zinc finger of GATA-1 reveals a specific binding face for the transcriptional co-factor FOG
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DOI：
10.1023/a:1008309602929
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发表时间：
1999-03-01
影响因子：
2.7
通讯作者：
Mackay, JP
中科院分区：
文献类型：
--
作者：
Kowalski, K;Czolij, R;Mackay, JP
锌指（ZnFs）通常被认为是DNA结合基序。然而，最近的一些报道暗示特定的ZnFs介导蛋白质-蛋白质相互作用。加塔-1（NF）的N-末端ZnF是这样一个指，已经显示与许多其他蛋白质相互作用，包括最近发现的转录辅因子FOG。在这里，我们解决了三维结构的NF在溶液中使用多维H-1/N-15 NMR光谱，我们使用H-1/N-15自旋关系测量，以调查其骨干动力学。该结构由两个扭曲的β-发夹和一个单一的α-螺旋组成，并且与鸡加塔-1的C-末端ZnF的结构相似。NF结构与其他C-4型锌结合基序，包括激素受体和LIM结构域的比较，也揭示了大量的结构同源性。最后，我们使用该结构来绘制通过诱变显示的对于FOG结合是必需的NF残基的空间位置，并证明这些残基都位于NE的单个面上。值得注意的是，该面与NE的推定DNA结合面很好地分离，这一观察结果提示NF的同时作用;即稳定加塔-1 DNA复合物和将FOG募集到加塔-1控制的启动子区域。
Zinc fingers (ZnFs) are generally regarded as DNA-binding motifs. However, a number of recent reports have implicated particular ZnFs in the mediation of protein-protein interactions. The N-terminal ZnF of GATA-1 (NF) is one such finger, having been shown to interact with a number of other proteins, including the recently discovered transcriptional co-factor FOG. Here we solve the three-dimensional structure of the NF in solution using multidimensional H-1/N-15 NMR spectroscopy, and we use H-1/N-15 spin relation measurements to investigate its backbone dynamics. The structure consists of two distorted beta-hairpins and a single alpha-helix, and is similar to that of the C-terminal ZnF of chicken GATA-1. Comparisons of the NF structure with those of other C-4-type zinc binding motifs, including hormone receptor and LIM domains, also reveal substantial structural homology. Finally, we use the structure to map the spatial locations of NF residues shown by mutagenesis to be essential for FOG binding, and demonstrate that these residues all lie on a single face of the NE Notably, this face is well removed from the putative DNA-binding face of the NE an observation which is suggestive of simultaneous roles for the NF; that is, stabilisation of GATA-1 DNA complexes and recruitment of FOG to GATA-1-controlled promoter regions.