Optimization of the efficiency of cross-linking PtII oligonucleotide phosphorothioate complexes to complementary oligonucleotides.
Optimization of the efficiency of cross-linking PtII oligonucleotide phosphorothioate complexes to complementary oligonucleotides.
复制标题
优化 PtII 寡核苷酸硫代磷酸酯复合物与互补寡核苷酸的交联效率。
DOI：
10.1093/nar/18.17.5163
复制
发表时间：
1990
影响因子：
14.9
通讯作者：
Orgel,LE
中科院分区：
文献类型：
--
作者：
Chu,BC;Orgel,LE
我们研究了PtII复合体将寡核苷酸的硫代产物与互补的DNA靶标进行交联的效率。A和G残基位于5‘-硫代硫酸酯基团下游的2-5个碱基是交联键的首选位置。当使用顺式二氯铂二胺(CisPtII)或氯化铂(K2PtCl4)时，用T残基取代目标这一部分的残基导致交联度大大降低。如果靶的敏感区域没有A和G残基，反式二氯铂二胺(TransPtII)与T残基形成交联键。含有内部硫代基团的低聚物也可以用TransPtII连接到它们的模板上，但不能用cisPtII或K2PtCl4连接。通过内部硫代基团进行的交联会比通过5‘-末端硫代磷酸盐进行的交联会效率低。内部硫代磷酸盐的SP型异构体比RPI型异构体更有效地交联。初步实验表明，与RNA靶标的交联效率将被证明与DNA靶标的效率相似。
We have investigated the efficiency with which PtIIcomplexes cross-link phosphorothloates of oligonucleotides to complementary DNA targets. The A and G residues 2–5 bases downstream from the 5′-phosphorothioate group are preferred sites for cross-linking. Replacement of residues in this part of the target by T residues results in greatly decreased cross-linking when cis platinum diammine dichloride (cisPtII) or potassium platinous chloride (K2PtCl4) are used. Trans platinum diammine dichloride (transPtII) forms cross-links with T residues if A and G residues are absent from the susceptible region of the target. Oligomers containing an internal phosphorothioate group can also be linked to their templates with transPtII, but not with cisPtIIor K2PtCl4. Cross-linking via an internal phosphorothioate group tends to be less efficient than cross-linking via a 5′-terminal phosphorothioate. The SPisomers of internal phosphorothioates are cross-linked more efficiently than the RPisomers. Preliminary experiments suggest that the efficiency of cross-linking to RNA targets will prove similar to that found for DNA targets.