Vorlícková M, Zimulová M, Kovanda J, Fojtík P, Kypr J
Institute of Biophysics, Academy of Sciences of the Czech Republic, Královopolská 135, CZ-612 65 Brno, Czech Republic.
Nucleic Acids Res. 1998 Jun 1;26(11):2679-85. doi: 10.1093/nar/26.11.2679.
We studied DNA dodecamers (CAG)4, (CCG)4, (CGG)4 and (CTG)4by CD spectroscopy and polyacrylamide gel electrophoresis. Each dodecamer adopted several ordered conformers which denatured in a cooperative way. Stability of the conformers depended on the dodecamer concentration, ionic strength, temperature and pH. The dodecamers, having a pyrimidine base in the triplet center, generated foldbacks at low ionic strength whose stem conformations were governed by the GC pairs. At high salt, (CCG)4 isomerized into a peculiar association of two strands. The association was also promoted by high oligonucleotide concentrations. No similar behavior was exhibited by (CTG)4. At low salt, (CGG)4 coexisted in two bimolecular conformers whose populations were strongly dependent on the ionic strength. In addition, (CGG)4 associated into a tetraplex at acidic pH. A tetraplex was even observed at neutral pH if the (CGG)4 concentration was sufficiently high. (CAG)4 was very stable in a monomolecular conformer similar to the known extremely stable foldback of the (GCGAAGC) heptamer. Nevertheless, even this very stable conformer disappeared if (CTG)4 was added to the solution of (CAG)4. Association of the complementary strands was also strongly preferred to the particular strand conformations by the other couple, (CCG)4 and (CGG)4.
我们通过圆二色光谱法和聚丙烯酰胺凝胶电泳研究了DNA十二聚体(CAG)4、(CCG)4、(CGG)4和(CTG)4。每个十二聚体都采用了几种有序构象,这些构象以协同方式变性。构象的稳定性取决于十二聚体浓度、离子强度、温度和pH值。在三联体中心含有嘧啶碱基的十二聚体在低离子强度下会产生回折,其茎部构象由GC对控制。在高盐条件下,(CCG)4异构化为两条链的特殊缔合形式。高寡核苷酸浓度也会促进这种缔合。(CTG)4没有表现出类似行为。在低盐条件下,(CGG)4以两种双分子构象共存,其数量强烈依赖于离子强度。此外,(CGG)4在酸性pH下缔合形成四链体。如果(CGG)4浓度足够高,在中性pH下甚至也能观察到四链体。(CAG)4在单分子构象中非常稳定,类似于已知的(GCGAAGC)七聚体极其稳定的回折结构。然而,如果将(CTG)4添加到(CAG)4溶液中,即使是这种非常稳定的构象也会消失。互补链的缔合也比另一对(CCG)4和(CGG)4的特定链构象更受青睐。