Mixed deoxyribo- and ribo-oligonucleotides with catalytic activity
Mixed deoxyribo- and ribo-oligonucleotides with catalytic activity
复制标题
具有催化活性的混合脱氧核糖和核糖寡核苷酸
作者：
J. Perreault;Taifeng Wu;B. Cousineau;K. K. Ogilvie;R. Cedergren
植物中类病毒和类病毒的RNA，以及蝾螈中一些串联重复DNA序列的RNA转录本，可以进行自催化切割，生成具有5''OH和2',3'-环磷酸末端的RNA14。这些催化RNA或核酶形成称为“ham-merhead”的茎环二级结构，其中催化（核酶）和底物序列靠近在一起（图1a）。使用寡核糖核苷酸可以很容易地制备具有催化活性的锤头状核酶模拟物57。因此，构建了锤头类似物，其中某些核糖核苷酸被不同的核糖核苷酸取代，以鉴定切割活性所需的共有残基并确定切割机制的细节810。但这些核糖核苷酸取代往往会通过改变取代位置的氢键和堆积电位来改变锤头的构象。我们现在构建了结构破坏较少的锤头类似物，其中缺少 2'-OH 基团的脱氧核糖核苷酸取代了核糖核苷酸。这些混合的 RNA-DNA 聚合物 11 是使用与 DNA 合成兼容的 RNA 化学合成策略合成的。对这些锤头类似物的裂解产物的分析证实了与底物中裂解位点相邻的2'-OH参与反应，并证明催化区域中的一些2'-OH基团强烈影响活性。结果还表明，产生三维结构的核酸型催化并不限于RNA。
THE RNA of viroids and virusoids in plants, and the RNA tran-scripts of some tandemly repeated DNA sequences in the newt, can undergo self-catalysed cleavage to generate RNA with 5'′OH and 2′,3′-cyclic-phosphate termini14. These catalytic RNAs, or ribozymes, form a stem-loop secondary structure called a 'ham-merhead' in which the catalytic (ribozyme) and substrate sequences are brought close together (Fig. la). Catalytically active mimics of hammerhead ribozymes can be readily made using oligoribo-nucleotides57. Consequently, hammerhead analogues in which certain ribonucleotides are replaced by different ones have been constructed both to identify consensus residues required for cleavage activity and to determine the details of the cleavage mechanism810. But these ribonucleotide-replacements tend to alter the conformation of the hammerhead by changing hydrogen-bonding and stacking potential at the position of substitition. We have now constructed structurally less-disrupted hammerhead analogues in which deoxyribonucleotides, which lack 2′-OH groups, are substituted for ribonucleotides. These mixed RNA-DNA polymers11 were synthesized using a strategy for the chemical synthesis of RNA that is compatible with DNA synthesis12. Analy-sis of the cleavage products of several of these hammerhead analogues confirms the involvement in the reaction of the 2′-OH adjacent to the cleavage site in the substrate, and demonstrates that some 2′-OH groups in the catalytic region strongly affect activity. The results also indicate that the three-dimensional struc-ture producing nucleic acid-type catalysis is not restricted to RNA.
含有共有 pribnow 启动子序列的合成 DNA-RNA 杂合十二聚体的高分辨率 NMR 研究：d(CGTTATAATGCG).r(CGCAUUAUAACG)。
影响因子：
2.9
作者：
Chou,SH;Flynn,P;Reid,B
通讯作者：
Reid,B
两种 RNA 催化的 RNA 裂解反应的表征。
DOI：
10.1101/sqb.1987.052.01.032
发表时间：
1987
期刊：
Cold Spring Harbor symposia on quantitative biology
影响因子：
--
作者：
Sampson,JR;Sullivan,FX;Behlen,LS;DiRenzo,AB;Uhlenbeck,OC
通讯作者：
Uhlenbeck,OC