期刊
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA
卷 107, 期 7, 页码 2872-2877出版社
NATL ACAD SCIENCES
DOI: 10.1073/pnas.0914869107
关键词
agmatine; decoding; RNA modification; tRNA; wobble pairing
资金
- National Institutes of Health [GM17151, GM22854, RR19900]
- National Science Foundation [CHE0602413, CHE0910751]
- University of Cincinnati
- U.S. Department of Energy [DE-FG36-08GO88055]
- Division Of Chemistry
- Direct For Mathematical & Physical Scien [0910751] Funding Source: National Science Foundation
Modification of the cytidine in the first anticodon position of the AUA decoding tRNA(Ile) (tRNA(2)(Ile)) of bacteria and archaea is essential for this tRNA to read the isoleucine codon AUA and to differentiate between AUA and the methionine codon AUG. To identify the modified cytidine in archaea, we have purified this tRNA species from Haloarcula marismortui, established its codon reading properties, used liquid chromatography-mass spectrometry (LC-MS) to map RNase A and T1 digestion products onto the tRNA, and used LC-MS/MS to sequence the oligonucleotides in RNase A digests. These analyses revealed that the modification of cytidine in the anticodon of tRNA(2)(Ile) adds 112 mass units to its molecular mass and makes the glycosidic bond unusually labile during mass spectral analyses. Accurate mass LC-MS and LC-MS/ MS analysis of total nucleoside digests of the tRNA(2)(Ile) demonstrated the absence in the modified cytidine of the C2-oxo group and its replacement by agmatine (decarboxy-arginine) through a secondary amine linkage. We propose the name agmatidine, abbreviation C+, for this modified cytidine. Agmatidine is also present in Methanococcus maripaludis tRNA(2)(Ile) and in Sulfolobus solfataricus total tRNA, indicating its probable occurrence in the AUA decoding tRNA(Ile) of euryarchaea and crenarchaea. The identification of agmatidine shows that bacteria and archaea have developed very similar strategies for reading the isoleucine codon AUA while discriminating against the methionine codon AUG.
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