Non-sequence-specific DNA binding by the FILAMENTOUS FLOWER protein from Arabidopsis thaliana is reduced by EDTA
Non-sequence-specific DNA binding by the FILAMENTOUS FLOWER protein from Arabidopsis thaliana is reduced by EDTA
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DOI：
10.1074/jbc.m108889200
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发表时间：
2002-04-05
影响因子：
4.8
通讯作者：
Okada, K
中科院分区：
文献类型：
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作者：
Kanaya, E;Nakajima, N;Okada, K
丝状花蛋白具有锌指结构域、疏水性区域、富含脯氨酸的区域和 HMG 盒状结构域。我们已经报道，锌指处的锌释放可能是由位置 56 处的非典型半胱氨酸残基促进的，并且 EDTA 引起结构变化并增强蛋白质的自组装（Kanaya, E.、Watanabe, K.、Nakajima, N.、Okada, K. 和 Shimura, Y. (2001) J. Bio. Chem. 276, 7383-7390)。为了进一步研究这个方面，我们检查了丝状花蛋白的 DNA 结合功能。凝胶阻滞实验表明，FILAMENTOUS FLOWER 蛋白与 DNA 的结合没有序列特异性。缺失分析表明，锌指结构域和疏水性区域不是必需的，但富含脯氨酸的区域和HMG盒状结构域对于FILAMENTOUS FLOWER蛋白的DNA结合是必不可少的。添加 EDTA 可以减少由锌指结构域和其余区域组成的蛋白质与 DNA 的结合。这一结果可能表明锌的释放、锌指结构域中可能发生的结构变化、分子间相互作用以及蛋白质的自组装与丝状花蛋白从DNA的解离有关。
The FILAMENTOUS FLOWER protein has a zinc finger domain, hydrophobic region, proline-rich region, and a HMG box-like domain. We have reported that zinc release at the zinc finger is probably facilitated by the non-canonical cysteine residue at position 56, and that EDTA causes the structural change and enhances the self-assembly of the protein (Kanaya, E., Watanabe, K., Nakajima, N., Okada, K., and Shimura, Y. (2001) J. Bio. Chem. 276, 7383-7390). To investigate this aspect further we examined the DNA binding function of the FILAMENTOUS FLOWER protein. Gel retardation experiments showed that the FILAMENTOUS FLOWER protein binds to DNA without sequence specificity. Deletion analyses suggested that the zinc finger domain and the hydrophobic region are not required but the proline-rich region and the HMG box-like domain are indispensable for the DNA binding by the FILAMENTOUS FLOWER protein. The DNA binding by the protein consisting of the zinc finger domain and the rest of the regions was reduced with the addition of EDTA. This result probably suggests that the zinc release, the structural change probably occurring in the zinc finger domain, the intermolecular interaction, and the self-assembly of the protein are related to the dissociation of the FILAMENTOUS FLOWER protein from DNA.