Inhibition of sequence-specific protein-DNA interaction and restriction endonuclease cleavage via triplex stabilization by poly(L-lysine)-graft-dextran copolymer.
Inhibition of sequence-specific protein-DNA interaction and restriction endonuclease cleavage via triplex stabilization by poly(L-lysine)-graft-dextran copolymer.
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通过聚（L-赖氨酸）-接枝-葡聚糖共聚物的三重稳定性抑制序列特异性蛋白质-DNA 相互作用和限制性内切核酸酶切割。
DOI：
10.1021/bm9900141
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发表时间：
2000
影响因子：
6.2
通讯作者：
A. Maruyama
中科院分区：
文献类型：
--
作者：
A. Ferdous;T. Akaike;A. Maruyama
哺乳动物基因启动子中的聚l -赖氨酸-接枝-葡聚糖共聚物在体外生理pH和离子条件下抑制HeLa细胞核蛋白的DNA结合活性以及限制性内切酶裂解。利用大鼠α -1 (I)胶原基因启动子的30-mer同嘌呤-同嘧啶延伸段（位于-170和-141 bp之间）进行的电泳迁移分析表明，该共聚物在其与DNA的电荷比范围内稳定了三联体DNA，并增强了三联体特异性抑制蛋白质-DNA相互作用。当启动子的三联体形成区域（位于-165和-146 bp之间）在质粒DNA的BamH1和Pst 1位点上进行工程设计时，共聚物介导的三联体稳定也显著地竞争了BamH1的内切酶活性。最后，该共聚物的三聚体稳定效率显著高于精胺和苯并[e]吡啶多酚。我们的研究结果表明，共聚物，不管目标双链的长度，稳定三链显著抑制蛋白质- dna相互作用和内切酶活性。由于在长天然双链的短区域内形成稳定的三聚体是赋予抗原策略治疗潜力的先决条件，长靶双链的三聚体稳定和核蛋白- dna相互作用的抑制可能打开共聚物在体内的适用性。
Triplex stabilization by poly(L-lysine)-graft-dextran copolymer within a mammalian gene promoter inhibits the DNA binding activity of nuclear proteins from HeLa cells as well as restriction endonuclease cleavage at physiological pH and ionic conditions in vitro. Electrophoretic mobility shift assays using a 30-mer homopurine-homopyrimidine stretch (located between -170 and -141 bp) of rat alpha 1 (I) collagen gene promoter reveal that the copolymer, at its wide range of charge ratio with DNA, stabilizes triplex DNA and enhances triplex-specific inhibition of the protein-DNA interaction. When the triplex-forming region (located between -165 and -146 bp) of the promoter is engineered at the Bam H1 and Pst 1 sites of a plasmid DNA, copolymer-mediated triplex stabilization also remarkably competes endonuclease activity of BamH1. Finally, the triplex-stabilizing efficiency of the copolymer is remarkably higher than that of spermine and benzo[e]pyridoindole. Our results indicate that the copolymer, regardless of the length of the target duplex, stabilizes triplexes for significant inhibition of protein-DNA interaction and endonuclease activity. Since stable triplex formation within a short region out of a long native duplex is a prerequisite to confer the therapeutic potential of antigene strategy, triplex stabilization on a long target duplex and inhibition of nuclear protein-DNA interaction may open the possible in vivo applicability of the copolymer.