Nazar R N, Wildeman A G
Nucleic Acids Res. 1981 Oct 24;9(20):5345-58. doi: 10.1093/nar/9.20.5345.
We have re-examined the nucleotide sequence of Vicia faba (broad bean) 5.8S rRNA using partial chemical degradation and a new approach to high temperature (65-80 degrees C) sequencing gels. The results indicate that the secondary structure was not completely disrupted in previous studies (Tanaka, Y., Dyer, T.A. and Brownlee, G.G. (1980) Nucleic Acid Res. 8, 1259-1272) and explain ambiguities between the nucleotide sequence and T1 ribonuclease digests. Despite this revision, estimates in the secondary structure suggest that this 5.8S rRNA differs from previously examined examples in two respects, more open conformations in both the "GC-rich" and "AU-rich" stems. The secondary structure was probed under a variety of ionic conditions using limited pancreatic and T1 ribonuclease digestion and rapid gel sequencing techniques. These studies and theoretical considerations generally supported the "burp gun" model previously proposed for all 5.8S rRNAs and were inconsistent with the recently suggested "cloverleaf" configuration. More importantly, they were also consistent with more open stem structures in this higher plant.
我们使用部分化学降解法和一种新的高温（65 - 80摄氏度）测序凝胶方法，重新检测了蚕豆5.8S rRNA的核苷酸序列。结果表明，在之前的研究中（Tanaka, Y., Dyer, T.A. 和Brownlee, G.G. (1980) Nucleic Acid Res. 8, 1259 - 1272），二级结构并未被完全破坏，这也解释了核苷酸序列与T1核糖核酸酶消化产物之间的模糊性。尽管有此修正，但二级结构的估计表明，这种5.8S rRNA在两个方面与之前检测的例子不同，即在“富含GC”和“富含AU”的茎中具有更开放的构象。我们在多种离子条件下，使用有限的胰核糖核酸酶和T1核糖核酸酶消化以及快速凝胶测序技术，对二级结构进行了探测。这些研究和理论考量总体上支持了之前为所有5.8S rRNAs提出的“冲锋枪”模型，与最近提出的“三叶草”构型不一致。更重要的是，它们也与这种高等植物中更开放的茎结构一致。
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