Solution structures of the DNA-binding domains of immune-related zinc-finger protein ZFAT
Solution structures of the DNA-binding domains of immune-related zinc-finger protein ZFAT
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DOI：
10.1007/s10969-015-9196-3
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发表时间：
2015-06-01
期刊：
影响因子：
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通讯作者：
Yokoyama, Shigeyuki
中科院分区：
文献类型：
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作者：
Tochio, Naoya;Umehara, Takashi;Yokoyama, Shigeyuki
ZFAT是一种转录调节因子，含有18个C2 H2型锌指和1个AT钩，参与自身免疫性甲状腺疾病、细胞凋亡和免疫相关细胞存活。我们通过核磁共振谱确定了13个ZFAT锌指（ZF）和从ZF 2到ZF 5区域的串联锌指的溶液结构。ZFAT具有8个不常见的凸出的含螺旋的锌指，并确定了其中6个结构（ZF 4，ZF 5，ZF 6，ZF 10，ZF 11和ZF 13）。ZFAT锌指蛋白中DNA结合位点的分布模式不同，表明N端和C端锌指蛋白具有不同的DNA序列偏好性。由于ZFAT具有三到五个连续的串联锌指，它们可以作为一个单元协同工作，我们还确定了两个串联排列的锌指结构，在ZF 2到ZF 4和ZF 3到ZF 5之间。我们的NMR光谱分析检测到ZF 4和ZF 5之间的相互作用，它们通过不常见的接头序列KKIK连接。ZF 4-ZF 5连接子抑制溶液中两个锌指之间的相对结构空间，与具有确定结构的其他连接子区域不同，表明ZF 4-ZF 5指间连接子参与ZFAT功能的调节。
ZFAT is a transcriptional regulator, containing eighteen C2H2-type zinc-fingers and one AT-hook, involved in autoimmune thyroid disease, apoptosis, and immune-related cell survival. We determined the solution structures of the thirteen individual ZFAT zinc-fingers (ZF) and the tandemly arrayed zinc-fingers in the regions from ZF2 to ZF5, by NMR spectroscopy. ZFAT has eight uncommon bulged-out helix-containing zinc-fingers, and six of their structures (ZF4, ZF5, ZF6, ZF10, ZF11, and ZF13) were determined. The distribution patterns of the putative DNA-binding surface residues are different among the ZFAT zinc-fingers, suggesting the distinct DNA sequence preferences of the N-terminal and C-terminal zinc-fingers. Since ZFAT has three to five consecutive tandem zinc-fingers, which may cooperatively function as a unit, we also determined two tandemly arrayed zinc-finger structures, between ZF2 to ZF4 and ZF3 to ZF5. Our NMR spectroscopic analysis detected the interaction between ZF4 and ZF5, which are connected by an uncommon linker sequence, KKIK. The ZF4-ZF5 linker restrained the relative structural space between the two zinc-fingers in solution, unlike the other linker regions with determined structures, suggesting the involvement of the ZF4-ZF5 interfinger linker in the regulation of ZFAT function.