Nazar R N, Sprott G D, Matheson A T, Van N T
Biochim Biophys Acta. 1978 Nov 21;521(1):288-94. doi: 10.1016/0005-2787(78)90271-x.
The secondary structure of 5-S rRNAs of Thermus aquaticus (an extreme thermophile), Bacillus stearothermophilus (a moderate thermophile) and Escherichia coli (a mesophile) was compared using thermal denaturation techniques under varying ionic conditions. At a low ionic strength (10 mM K+), the Tm of T. aquaticus 5-S RNA differed by only 1 degrees C from that of E. coli RNA and the molecule was fully denatured well below the optimum growth temperature of the thermophile. The internal Na+, K+ and Mg2+ concentrations of T. aquaticus cells were determined to be 91 mM, 130 mM and 59 mM, respectively. Under these salt conditions, T. aquaticus 5-S RNA was significantly more stable than E. coli RNA and the 5-S RNA from B. stearothermophilus was intermediate as is its optimum growth temperature. The results suggest that the thermostability of macromolecules from thermophilic organisms may be specially dependent on the internal salt concentration. Furthermore, under these salt conditions, most of the secondary structure of the RNA remained stable at the optimum growth temperatures suggesting that ribosomal RNAs of thermophilic organisms contribute more to the thermostability of the ribosome than previously thought.
利用热变性技术,在不同离子条件下比较了嗜热栖热菌(一种极端嗜热菌)、嗜热脂肪芽孢杆菌(一种中度嗜热菌)和大肠杆菌(一种嗜温菌)5 - S rRNA的二级结构。在低离子强度(10 mM K⁺)下,嗜热栖热菌5 - S RNA的解链温度与大肠杆菌RNA的解链温度仅相差1℃,并且该分子在远低于嗜热菌最佳生长温度时就完全变性。测定嗜热栖热菌细胞内的Na⁺、K⁺和Mg²⁺浓度分别为91 mM、130 mM和59 mM。在这些盐条件下,嗜热栖热菌5 - S RNA比大肠杆菌RNA显著更稳定,而嗜热脂肪芽孢杆菌的5 - S RNA则处于中间状态,其最佳生长温度也是如此。结果表明,嗜热生物大分子的热稳定性可能特别依赖于细胞内盐浓度。此外,在这些盐条件下,RNA的大部分二级结构在最佳生长温度下仍保持稳定,这表明嗜热生物的核糖体RNA对核糖体热稳定性的贡献比以前认为的更大。