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5S rRNA modification in the hyperthermophilic archaea Sulfolobus solfataricus and Pyrodictium occultum
Author(s) -
Bruenger Eveline,
Kowalak Jeffrey A.,
Kuchino Yoshiyuki,
McCloskey James A.,
Mizushima Hiroshi,
Stetter Karl O.,
Grain Pamela F.
Publication year - 1993
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.7.1.8422966
Subject(s) - sulfolobus solfataricus , sulfolobus , thermophile , ribosomal rna , archaea , biochemistry , chemistry , oligonucleotide , biology , enzyme , gene
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, N 4 ‐acetylcytidine (ac 4 C) and N 4 ‐acetyl‐2′‐ O ‐methylcytidine (ac 4 Cm). Oligonucleotides were generated from P. occultum 5S rRNA by RNase T 1 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 ac 4 Cm to be located at position 35, in the nonanucleotide CAA‐CACC[ac 4 Cm]G, and the ac 4 C 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 ac 4 C and ac 4 Cm on thermal stabilization of 5S rRNA in thermophiles are discussed.— Bruenger, E., Kowalak, J. A., Kuchino, Y., McCloskey, J. A., Mizushima, H., Stetter, K. O., Grain, P. F. 5S rRNA modification in the hyperthermophilic archaea Sulfolobus solfataricus and Pyrodictium occultum . FASEB J. 7: 196‐200; 1993.