Ohta S, Maruyama S, Nitta K, Sugai S
Nucleic Acids Res. 1983 May 25;11(10):3363-73. doi: 10.1093/nar/11.10.3363.
Equilibrium and kinetics of thermal melting of yeast 5S ribosomal RNA in aqueous NaCl with or without Mg2+ were investigated by differential thermal melting and temperature jump methods. Two peaks (1 and 2) and a shoulder were observed in each of melting curves at ionic strength I=0.002-0.5 and linearity between each of melting temperatures T1m and T2m and log I was found at I=0.01-0.5 in the Mg2+-free solution. The local structures were found to be stabilized considerably by Mg2+. The temperature jump measurements gave the kinetic melting curve of the structure 1 at I=0.03 without Mg2+ or with 0.5 mM Mg2+. The kinetic Tm coincided well with the corresponding static Tm. For the structure 1, various parameters were calculated from the kinetic data, which indicated a double helical character of the structure 1. In terms of the values of Tm, G-C content, and enthalpy change of the transition of the structure 1 or 2, appropriateness of each of the secondary structure models of eukaryotic 5S RNA proposed previously was discussed.
采用差示热熔解和温度跃升方法研究了在有或无Mg2+存在的情况下,酵母5S核糖体RNA在NaCl水溶液中的热熔解平衡和动力学。在离子强度I = 0.002 - 0.5时,每条熔解曲线中均观察到两个峰(1和2)及一个肩峰,且在无Mg2+的溶液中,于I = 0.01 - 0.5时发现每个熔解温度T1m和T2m与log I之间呈线性关系。发现局部结构因Mg2+而得到显著稳定。温度跃升测量给出了在I = 0.03、无Mg2+或含有0.5 mM Mg2+时结构1的动力学熔解曲线。动力学熔解温度(Tm)与相应的静态Tm吻合良好。对于结构1,从动力学数据计算出各种参数,这表明结构1具有双螺旋特征。根据结构1或2的Tm值、G - C含量及转变的焓变,讨论了先前提出的真核生物5S RNA各二级结构模型的适宜性。