Khomyakova E B, Gousset H, Liquier J, Huynh-Dinh T, Gouyette C, Takahashi M, Florentiev V L, Taillandier E
Laboratoire de Spectroscopie Biomoléculaire, URA CNRS 1430, Université Paris Nord, 74 rue Marcel Cachin, F-93017 Bobigny Cedex, France.
Nucleic Acids Res. 2000 Sep 15;28(18):3511-6. doi: 10.1093/nar/28.18.3511.
We present evidence of formation of an intramolecular parallel triple helix with TA.T and GG.C base triplets (where * represents the hydrogen bonding interaction between the third strand and the duplex while. represents the Watson-Crick interactions which stabilize the duplex). The third GT strand, containing seven GpT/TpG steps, targets the polypurine sequence 5'-AGG-AGG-GAG-GAG-3'. The triple helix is obtained by the folding back twice of a 36mer, formed by three dodecamers tethered by hydroxyalkyl linkers (-L-). Due to the design of the oligonucleotide, the third strand orientation is parallel with respect to the polypurine strand. Triple helical formation has been studied in concentration conditions in which native gel electrophoresis experiments showed the absence of intermolecular structures. Circular dichroism (CD) and UV spectroscopy have been used to evidence the triplex structure. A CD spectrum characteristic of triple helical formation as well as biphasic UV and CD melting curves have been obtained in high ionic strength NaCl solutions in the presence of Zn(2+) ions. Specific interactions with Zn(2+) ions in low water activity conditions are necessary to stabilize the parallel triplex.
我们展示了由TA.T和GG.C碱基三联体形成分子内平行三链螺旋的证据(其中*表示第三链与双链体之间的氢键相互作用,而.表示稳定双链体的沃森-克里克相互作用)。第三条GT链包含七个GpT/TpG步移,靶向多嘌呤序列5'-AGG-AGG-GAG-GAG-3'。三链螺旋是通过由羟烷基接头(-L-)连接的三个十二聚体形成的36聚体两次回折得到的。由于寡核苷酸的设计,第三链的方向与多嘌呤链平行。在天然凝胶电泳实验表明不存在分子间结构的浓度条件下研究了三链螺旋的形成。圆二色性(CD)和紫外光谱已被用于证明三链体结构。在高离子强度的NaCl溶液中,在Zn(2+)离子存在的情况下,获得了三链螺旋形成的特征性CD光谱以及双相紫外和CD熔解曲线。在低水活度条件下与Zn(2+)离子的特异性相互作用对于稳定平行三链体是必要的。