DNA recognition by Escherichia coli CbpA protein requires a conserved arginine-minor-groove interaction.
DNA recognition by Escherichia coli CbpA protein requires a conserved arginine-minor-groove interaction.
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DOI：
10.1093/nar/gkv012
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发表时间：
2015-02-27
影响因子：
14.9
通讯作者：
Grainger DC
中科院分区：
文献类型：
--
作者：
Chintakayala K;Sellars LE;Singh SS;Shahapure R;Westerlaken I;Meyer AS;Dame RT;Grainger DC
弯曲DNA结合蛋白A（covered DNA binding protein A，CbpA）是一种存在于γ-变形菌中的DNA结合蛋白。在大肠杆菌中研究得最好的是，CbpA在饥饿期间积累到每个细胞> 2500个拷贝，并与DNA形成聚集体。然而，DNA结合的分子基础是未知的; CbpA缺乏在其他细菌DNA结合蛋白中发现的基序。在这里，我们使用遗传学和生物化学的结合来阐明CbpA识别DNA的机制。我们发现，CbpA与DNA小沟相互作用。这种相互作用需要高度保守的精氨酸侧链。用丙氨酸取代该残基R116，特异性地破坏CbpA及其来自其它细菌的同系物与DNA的结合，同时不影响其它CbpA活性。当DNA结合被否定时，CbpA的细胞内分布显著改变。因此，我们提供了DNA结合和细胞中CbpA行为之间的直接联系。
Curved DNA binding protein A (CbpA) is a co-chaperone and nucleoid associated DNA binding protein conserved in most γ-proteobacteria. Best studied in Escherichia coli, CbpA accumulates to >2500 copies per cell during periods of starvation and forms aggregates with DNA. However, the molecular basis for DNA binding is unknown; CbpA lacks motifs found in other bacterial DNA binding proteins. Here, we have used a combination of genetics and biochemistry to elucidate the mechanism of DNA recognition by CbpA. We show that CbpA interacts with the DNA minor groove. This interaction requires a highly conserved arginine side chain. Substitution of this residue, R116, with alanine, specifically disrupts DNA binding by CbpA, and its homologues from other bacteria, whilst not affecting other CbpA activities. The intracellular distribution of CbpA alters dramatically when DNA binding is negated. Hence, we provide a direct link between DNA binding and the behaviour of CbpA in cells.
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影响因子：
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作者：
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通讯作者：
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Grainger DC
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