A putative DNA-binding domain in the NUCKS protein
A putative DNA-binding domain in the NUCKS protein
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DOI：
10.1016/s0003-9861(02)00513-1
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发表时间：
2002-11-15
影响因子：
3.9
通讯作者：
Ostvold, AC
中科院分区：
文献类型：
--
作者：
Grundt, K;Skjeldal, L;Ostvold, AC
我们研究了一种由NUCKS衍生的、含有扩展的GRP基序的合成肽的DNA结合特性。该肽可与随机序列DNA结合，但不优先与聚(da-dt)结合。具有相同氨基酸组成但具有随机序列的合成肽不与DNA结合，这表明DNA结合区的结构在与DNA的相互作用中起着关键作用。用核磁共振和图形模拟对合成肽的结构进行了研究。结果表明，DNA结合肽在pH为5.5的磷酸盐缓冲液中形成一个α螺旋。对接结果表明，这种小的螺旋多肽几乎完全适合DNA的主槽，有可能有四个碱性残基与DNA的磷酸骨架相互作用。CDK1的一个共同的磷酸化位点位于DNA结合肽的N端。一旦该位点被磷酸化，与DNA的结合就被完全禁止。免疫荧光实验表明，在细胞周期间期，NUCKS定位于增殖细胞的胞核中，而在有丝分裂细胞中则分布于胞浆中。(C)2002年埃尔塞维尔科学公司(美国)。版权所有。
We have studied the DNA-binding properties of a NUCKS-derived, synthetic peptide containing an extended GRP motif. This peptide binds to random-sequence DNA, but did not bind preferentially to poly(dA-dT). A synthetic peptide with the same amino acid composition but with a random sequence did not bind to DNA, suggesting that the structure of the DNA-binding domain plays a pivotal role in the interaction with DNA. NMR and graphic modeling were employed to investigate the structure of the synthetic peptide. It was shown that the DNA-binding peptide constituted an alpha helix in phosphate buffer at pH 5.5. Docking results indicated an almost perfect fit for this small, helical peptide into the major groove of DNA with the possibility of four basic residues interacting with the phosphate backbone of DNA. One consensus site for phosphorylation by Cdk1 is located in the N-terminal end of the DNA-binding peptide. Upon phosphorylation of this site, the binding to DNA was completely prohibited. Immunofluorescence experiments showed that NUCKS was located in the nuclei in proliferating cells in interphase of the cell cycle, but was distributed throughout the cytoplasm in mitotic cells. (C) 2002 Elsevier Science (USA). All rights reserved.