Solution Structure of Gfi-1 Zinc Domain Bound to Consensus DNA
Solution Structure of Gfi-1 Zinc Domain Bound to Consensus DNA
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DOI：
10.1016/j.jmb.2010.02.006
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发表时间：
2010-04-09
影响因子：
5.6
通讯作者：
Wu, Zhengrong
中科院分区：
文献类型：
--
作者：
Lee, Soojin;Doddapaneni, Kiran;Wu, Zhengrong
GFI-1是一种重要的转录抑制因子，在造血过程中对细胞的增殖和分化起着重要的调控作用。最近，这种蛋白质也被证明能够限制造血干细胞的增殖，这一过程似乎对造血干细胞的长期能力至关重要。这两个看似相反的监管结果很可能来自于它与各种细胞合作伙伴的互动。这种相互作用可以直接影响GFI-1通过其DNA结合的锌指结构域识别的基因。在这项工作中，我们报道了用多维核磁共振方法测定GFI-1锌指3-5与16聚体共有DNA的络合物的结构。与基于甲基化干扰实验的次要凹槽结合模型不同，我们的结构清楚地表明GFI-1锌指3-5结合到靶DNA的主要凹槽中，这使人想起典型的C2H2锌指结构域。第四和第五个锌指通过在Asn382、Gln379和Asp354的侧链与不变腺嘌呤和胞嘧啶的碱基之间形成碱基特异性氢键来识别AATC核心序列。总体而言，目前的工作为DNA结合特异性的结构决定因素提供了有价值的见解，特别是对于在任何其他结构的锌指-DNA复合体中未观察到的TCA三联体，以及导致急性髓系白血病的自然发生突变的分子基础。(C)2010爱思唯尔有限公司。保留所有权利。GFI-1是一种关键的转录抑制因子，用于精确调控造血过程中的细胞增殖和分化。最近，这种蛋白质也被证明能够限制造血干细胞的增殖，这一过程似乎对造血干细胞的长期能力至关重要。这两个看似相反的监管结果很可能来自于它与各种细胞合作伙伴的互动。这种相互作用可以直接影响GFI-1通过其DNA结合的锌指结构域识别的基因。在这项工作中，我们报道了用多维核磁共振方法测定GFI-1锌指3-5与16聚体共有DNA的络合物的结构。与基于甲基化干扰实验的次要凹槽结合模型不同，我们的结构清楚地表明GFI-1锌指3-5结合到靶DNA的主要凹槽中，这使人想起典型的C2H2锌指结构域。第四和第五个锌指通过在Asn382、Gln379和Asp354的侧链与不变腺嘌呤和胞嘧啶的碱基之间形成碱基特异性氢键来识别AATC核心序列。总体而言，目前的工作为DNA结合特异性的结构决定因素提供了有价值的见解，特别是对于在任何其他结构的锌指-DNA复合体中未观察到的TCA三联体，以及导致急性髓系白血病的自然发生突变的分子基础。(C)2010爱思唯尔有限公司。保留所有权利。
Gfi-1 is a crucial transcriptional repressor for the precise regulation of cell proliferation and differentiation in hematopoiesis. Recently, this protein has also been demonstrated to be capable of restricting the proliferation of hematopoietic stem cells, a process that appears to be vital for the long-term competency of hematopoietic stem cells. These two seemingly opposite outcomes of regulation are likely to arise from its interactions with a variety of cellular partners. Such interactions can directly affect the genes that Gfi-1 recognizes through its DNA binding zinc-finger domain. In this work, we report the determination of the solution structure of Gfi-1 zinc fingers 3-5 in complex with a 16-mer consensus DNA using multidimensional NMR method. Unlike a proposed minor-groove binding model based on methylation interference experiments, our structure clearly shows that Gfi-1 zinc fingers 3-5 bind into the major groove of the target DNA reminiscent of canonical C2H2 zinc-finger domains. The fourth and fifth zinc fingers recognize the AATC core sequence by forming base-specific hydrogen bonds between the side chains of Asn382, Gln379, and Asp354 and the bases of the invariant adenines and cytosine. Overall, the current work provides valuable insight into the structural determinants for DNA binding specificity, in particular for the TCA triplet that has not been observed in any other structures of zinc finger-DNA complexes, as well as molecular rationales for a naturally occurring mutation that causes acute myeloid leukemia. (C) 2010 Elsevier Ltd. All rights reserved.Gfi-1 is a crucial transcriptional repressor for the precise regulation of cell proliferation and differentiation in hematopoiesis. Recently, this protein has also been demonstrated to be capable of restricting the proliferation of hematopoietic stem cells, a process that appears to be vital for the long-term competency of hematopoietic stem cells. These two seemingly opposite outcomes of regulation are likely to arise from its interactions with a variety of cellular partners. Such interactions can directly affect the genes that Gfi-1 recognizes through its DNA binding zinc-finger domain. In this work, we report the determination of the solution structure of Gfi-1 zinc fingers 3-5 in complex with a 16-mer consensus DNA using multidimensional NMR method. Unlike a proposed minor-groove binding model based on methylation interference experiments, our structure clearly shows that Gfi-1 zinc fingers 3-5 bind into the major groove of the target DNA reminiscent of canonical C2H2 zinc-finger domains. The fourth and fifth zinc fingers recognize the AATC core sequence by forming base-specific hydrogen bonds between the side chains of Asn382, Gln379, and Asp354 and the bases of the invariant adenines and cytosine. Overall, the current work provides valuable insight into the structural determinants for DNA binding specificity, in particular for the TCA triplet that has not been observed in any other structures of zinc finger-DNA complexes, as well as molecular rationales for a naturally occurring mutation that causes acute myeloid leukemia. (C) 2010 Elsevier Ltd. All rights reserved.