Wildeman A G, Nazar R N
J Biol Chem. 1982 Oct 10;257(19):11395-404.
The cytoplasmic ribosomes of the thermophilic fungus Thermomyces lanuginosus contain two types of 5 S RNA. The nucleotide sequence for approximately 80% of the molecules is (pp)pA-C-A-U-G-C-G-A-C-C-A-U-A-G-G-G-U-G-U-G-G-A-A-A-A-C-A-G-G-G-C-U-U-C-C-C-G-U-C-C-G-C-U-C-A-G-C-C-G-U-A-C-U-U-A-A-G-C-C-A-C-A-C-G-C-C-G-G-C-U-G-G-U-U-A-G-U-A-G-U-U-G-G-G-U-G-G-G-U-G-A-C-C-A-C-C-A-G-C-G-A-A-U-C-C-C-A-G-C-U-G-U-U-G-C-A-U-G-UOH. The remainder contains two nucleotide substitutions, C19 and G60, which preserve base complementarity. The secondary structure was probed using partial T1, pancreatic, and S1 nuclease digestion under a variety of ionic and temperature conditions and fragments were analyzed by rapid gel sequencing techniques. The results support the Y-shaped secondary structure model originally proposed by Nishikawa, K., and Takemura, S. (1974) FEBS Lett. 40, 106-109, for eukaryotic 5 S RNAs. When the thermal denaturation profile was compared with that of the yeast 5 S RNA, the thermophilic RNA exhibited not only a higher Tm but also an unusual decline in absorbency at moderate temperatures. This suggests that a functionally important structure may be maintained only at higher temperatures.
嗜热真菌嗜热栖热菌的细胞质核糖体含有两种类型的5S RNA。约80%的分子的核苷酸序列为(pp)pA - C - A - U - G - C - G - A - C - C - A - U - A - G - G - G - U - G - U - G - G - A - A - A - A - C - A - G - G - G - C - U - U - C - C - C - G - U - C - C - G - C - U - C - A - G - C - C - G - U - A - C - U - U - A - G - C - C - A - C - A - C - G - C - C - G - G - C - U - G - G - U - U - A - G - U - A - G - U - U - G - G - G - U - G - G - G - U - G - A - C - C - A - C - C - A - G - C - G - A - A - U - C - C - C - A - G - C - U - G - U - U - G - C - A - U - G - UOH。其余部分含有两个核苷酸取代,即C19和G60,它们保持了碱基互补性。在各种离子和温度条件下,使用部分T1核酸酶、胰核酸酶和S1核酸酶消化来探测二级结构,并通过快速凝胶测序技术分析片段。结果支持了西川K.和武村S.(1974年)在《欧洲生物化学学会联合会快报》40卷,第106 - 109页中最初提出的真核生物5S RNA的Y形二级结构模型。当将热变性曲线与酵母5S RNA的热变性曲线进行比较时,嗜热RNA不仅表现出更高的解链温度,而且在中等温度下吸光度出现异常下降。这表明一个功能重要的结构可能仅在较高温度下得以维持。