NUCLEASE ACTIVITY OF 1,10-PHENANTHROLINE-COPPER ION

1，10-菲咯啉-铜（OP-Cu）对RNA和DNA的核酸酶活性 通过对核糖部分的氧化攻击。自由配位络合物是 可用于研究核酸结构/功能关系。 连接到载体配体，OP-Cu允许合成核酸酶， 设计的特殊性。 将开发用于基因组作图的靶向核酸酶活性， 涉及a）细菌阻遏物的基因工程衍生物， cro蛋白;和B）使用经修饰的RNA形成R环 1，10-菲咯啉这两种方法将能够在任何选择的切割 标记基因序列基因特异性抑制剂的合成 转录将通过将OP-Cu连接到寡核苷酸来实现a） 与转录起始位点的单链区域互补; 或B）能够与聚嘌呤/聚嘧啶形成形式三螺旋 真核基因侧翼的片段。 核糖核酸酶的化学机制和二级结构特异性 将测定OP-Cu的活性。沿着光活化铀酰 乙酸，其作为RNA-蛋白质足迹试剂的潜力 互动将被探索。三重螺旋形成， 双链RNA和脱氧寡核苷酸化学衍生， 将研究OP-Cu。通过分析目标的断裂模式 RNA，三螺旋形成的严格性，极性和稳定性将 下定决心。这种相互作用为设计 反义试剂在RNA水平阻断基因表达。

The nuclease activity of 1,10-phenanthroline-Cu (OP-Cu) nicks RNA and DNA by oxidative attack on the ribose moiety. The free coordination complex is useful for investigating nucleic acid structure/function relationships. Linked to carrier ligands, OP-Cu allows synthesis of nucleases with designed specificities. Targeted nuclease activities for genomic mapping will be developed which involve a) genetically engineered derivatives of the bacterial repressor cro protein; and b) R-loop formation using RNAs modified with 1,10-phenanthroline. Both methods will be capable of scission at any chosen marker gene sequence. The synthesis of gene specific inhibitors of transcription will be achieved by linking OP-Cu to oligonucleotides a) complementary to single stranded regions of transcription initiation sites; or b) capable of formal, triple helices with polypurine/polypyrimidine tracts which flank eucaryotic genes. The chemical mechanism and secondary structure specificity of the RNase activity of OP-Cu will be determined. Along with light-activated uranyl acetate, its potential as a footprinting reagent for RNA-protein interactions will be explored. Triple helix formation between double-stranded RNA and deoxyoligonucleotides chemically derivatized with OP-Cu will be investigated. By analyzing the scission pattern of the target RNA, the stringency, polarity, and stability of triple helix formation will be determined. This interaction provides a new approach for the design of antisense reagents to block gene expression at the level of RNA.