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首页> 外文期刊>Molecular and Cellular Biology >Importance of a flanking AT-rich region in target site recognition by the GC box-binding zinc finger protein MIG1.
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Importance of a flanking AT-rich region in target site recognition by the GC box-binding zinc finger protein MIG1.

机译：GC框结合锌指蛋白MIG1的侧翼富含AT的区域在目标位点识别中的重要性。

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摘要

MIG1 is a zinc finger protein that mediates glucose repression in the yeast Saccharomyces cerevisiae. MIG1 is related to the mammalian Krox/Egr, Wilms' tumor, and Sp1 finger proteins. It has two fingers and binds to a GCGGGG motif that resembles the GC boxes recognized by these mammalian proteins. We have performed a complete saturation mutagenesis of a natural MIG1 site in order to elucidate its binding specificity. We found that only three mutations within the GC box retain the ability to bind MIG1: G1 to C, C2 to T, and G5 to A. This result is consistent with current models for zinc finger-DNA binding, which assume that the sequence specificity is determined by base triplet recognition within the GC box. Surprisingly, we found that an AT-rich region 5' to the GC box also is important for MIG1 binding. This AT box is present in all natural MIG1 sites, and it is protected by MIG1 in DNase I footprints. However, the AT box differs from the GC box in that no single base within it is essential for binding. Instead, the AT-rich nature of this sequence seems to be crucial. The fact that AT-rich sequences are known to increase DNA flexibility prompted us to test whether MIG1 bends DNA. We found that binding of MIG1 is associated with bending within the AT box. We conclude that DNA binding by a simple zinc finger protein such as MIG1 can involve both recognition of the GC box and flanking sequence preferences that may reflect local DNA bendability.
机译：MIG1是一种锌指蛋白，可在酿酒酵母中介导葡萄糖抑制。 MIG1与哺乳动物的Krox / Egr，Wilms肿瘤和Sp1手指蛋白有关。它有两个手指，并与类似于这些哺乳动物蛋白识别的GC盒的GCGGGG基序结合。我们已经对天然MIG1位点进行了完全的饱和诱变，以阐明其结合特异性。我们发现，GC框内只有三个突变保留了将MIG1结合的能力：G1到C，C2到T和G5到A。此结果与当前的锌指DNA结合模型一致，这些模型假定序列特异性由GC框内的基本三元组识别确定。出人意料的是，我们发现GC框的5'富AT区对于MIG1结合也很重要。该AT盒存在于所有自然MIG1站点中，并且在DNase I足迹中受到MIG1的保护。但是，AT盒与GC盒的不同之处在于，其中没有单个碱基对于绑定必不可少。取而代之的是，该序列的AT丰富特性似乎至关重要。已知富含AT的序列可以增加DNA的灵活性，这一事实促使我们测试MIG1是否会弯曲DNA。我们发现，MIG1的绑定与AT盒内的弯曲相关。我们得出的结论是，简单的锌指蛋白（例如MIG1）与DNA的结合可能涉及对GC盒的识别以及可能反映局部DNA易弯曲性的侧翼序列偏好。

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