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首页> 外文期刊>Molecular and Cellular Biology >Characterization of the DNA-binding properties of the myeloid zinc finger protein MZF1: two independent DNA-binding domains recognize two DNA consensus sequences with a common G-rich core.
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Characterization of the DNA-binding properties of the myeloid zinc finger protein MZF1: two independent DNA-binding domains recognize two DNA consensus sequences with a common G-rich core.

机译：髓系锌指蛋白MZF1的DNA结合特性的表征：两个独立的DNA结合域识别具有共同的富含G核的两个DNA共有序列。

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The myeloid zinc finger gene 1, MZF1, encodes a transcription factor which is expressed in hematopoietic progenitor cells that are committed to myeloid lineage differentiation. MZF1 contains 13 C2H2 zinc fingers arranged in two domains which are separated by a short glycine- and proline-rich sequence. The first domain consists of zinc fingers 1 to 4, and the second domain is formed by zinc fingers 5 to 13. We have determined that both sets of zinc finger domains bind DNA. Purified, recombinant MZF1 proteins containing either the first set of zinc fingers or the second set were prepared and used to affinity select DNA sequences from a library of degenerate oligonucleotides by using successive rounds of gel shift followed by PCR amplification. Surprisingly, both DNA-binding domains of MZF1 selected similar DNA-binding consensus sequences containing a core of four or five guanine residues, reminiscent of an NF-kappa B half-site: 1-4, 5'-AGTGGGGA-3'; 5-13, 5'-CGGGnGAGGGGGAA-3'. The full-length MZF1 protein containing both sets of zinc finger DNA-binding domains recognizes synthetic oligonucleotides containing either the 1-4 or 5-13 consensus binding sites in gel shift assays. Thus, we have identified the core DNA consensus binding sites for each of the two DNA-binding domains of a myeloid-specific zinc finger transcription factor. Identification of these DNA-binding sites will allow us to identify target genes regulated by MZF1 and to assess the role of MZF1 as a transcriptional regulator of hematopoiesis.
机译：骨髓锌指基因1（MZF1）编码一个转录因子，该转录因子在致力于骨髓谱系分化的造血祖细胞中表达。 MZF1包含13个C2H2锌指，排列在两个结构域中，这些结构域由短的富含甘氨酸和脯氨酸的序列隔开。第一个域由锌指1至4组成，第二个域由锌指5至13形成。我们已经确定两组锌指域均结合DNA。制备了包含第一组锌指或第二组锌的纯化的重组MZF1蛋白，并用于通过使用连续几轮的凝胶移位和PCR扩增从简并寡核苷酸文库中亲和选择DNA序列。出人意料的是，MZF1的两个DNA结合结构域都选择了相似的DNA结合共有序列，其含有四个或五个鸟嘌呤残基的核心，让人联想到NF-κB半位点：1-4、5'-AGTGGGGA-3'; 5-13，5'-CGGGnGAGGGGGAA-3'。包含两组锌指DNA结合结构域的全长MZF1蛋白在凝胶位移测定中识别包含1-4或5-13共有结合位点的合成寡核苷酸。因此，我们已经确定了髓样特异性锌指转录因子的两个DNA结合域的核心DNA共有结合位点。这些DNA结合位点的鉴定将使我们能够鉴定由MZF1调控的靶基因，并评估MZF1作为造血功能的转录调控因子的作用。

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