5S RIBOSOMAL-RNA MODIFICATION IN THE HYPERTHERMOPHILIC ARCHAEA SULFOLOBUS-SOLFATARICUS AND PYRODICTIUM-OCCULTUM
5S RIBOSOMAL-RNA MODIFICATION IN THE HYPERTHERMOPHILIC ARCHAEA SULFOLOBUS-SOLFATARICUS AND PYRODICTIUM-OCCULTUM
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DOI：
10.1096/fasebj.7.1.8422966
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发表时间：
1993-01-01
期刊：
影响因子：
4.8
通讯作者：
CRAIN, PF
中科院分区：
文献类型：
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作者：
BRUENGER, E;KOWALAK, JA;CRAIN, PF
将硫磺硫化叶菌（Sulfolobus solfataricus）和隐热解菌（Pyrodicium occultum）的5S rRNA消化成核苷，并使用直接组合的HPLC/质谱法进行分析。P. occultum 5S rRNA含有两种修饰的核苷种类，N4-乙酰基胞苷（ac 4C）和N4-乙酰基-2 '-O-甲基胞苷（ac 4Cm）。通过RNase T1水解从P. occultum 5S rRNA产生寡核苷酸，并使用电喷雾质谱法测定其分子量，并与从P. occultum 5S RNA基因序列预测的分子量进行比较。预期和观察到的分子量之间的质量偏差允许ac 4Cm位于第35位，在九核苷酸CAA-CACC[ac 4Cm]G中，并且ac 4C在两个（C，U）G三核苷酸中的一个或两个中。2 '-O-甲基胞苷在S. solfataricus 5S rRNA，证实了早期在类似序列位置的暂定分配（斯塔尔，D.一、Luehrsen，K. R.，韦斯角，澳-地R.，和Pace，N. R.（1981）Nucleic Acids Res.，第9卷，第6129-6137; Dams，E.，隆代山口Cammarano，P.，Vandenberghe，A.，和De Wachter，R.等（1983）Nucleic Acids Res.第11卷，第111页。4667-4676）。ac 4C和ac 4Cm的存在下对5S rRNA在嗜热菌中的热稳定性的潜在影响进行了讨论。
The 5S rRNAs from Sulfolobus solfataricus and Pyrodictium occultum were digested to nucleosides and analyzed using directly-combined HPLC/mass spectrometry. P. occultum 5S rRNA contains two modified nucleoside species, N4-acetylcytidine (ac4C) and N4-acetyl-2'-O-methylcytidine (ac4Cm). Oligonucleotides were generated from P. occultum 5S rRNA by RNase T1 hydrolysis, and their molecular weights were determined using electrospray mass spectrometry and compared with those predicted from the P. occultum 5S RNA gene sequence. Deviation in mass between expected and observed molecular weights permitted ac4Cm to be located at position 35, in the nonanucleotide CAA-CACC[ac4Cm]G, and the ac4C in one or both of two (C,U)G trinucleotides. 2'-O-Methylcytidine is unambiguously characterized in S. solfataricus 5S rRNA, confirming earlier tentative assignments at the analogous sequence position (Stahl, D. A., Luehrsen, K. R., Woese, C. R., and Pace, N. R. (1981) Nucleic Acids Res., Vol. 9, pp. 6129-6137; Dams, E., Londei, P.. Cammarano, P., Vandenberghe, A., and De Wachter, R. (1983) Nucleic Acids Res. Vol. 11, pp. 4667-4676). Potential effects of the presence of ac4C and ac4Cm on thermal stabilization of 5S rRNA in thermophiles are discussed.