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- Title
Transfer RNA Methyltransferases from Thermoplasma acidophilum, a Thermoacidophilic Archaeon.
- Authors
Takuya Kawamura; Ryou Anraku; Takahiro Hasegawa; Chie Tomikawa; Hiroyuki Hori
- Abstract
We investigated tRNA methyltransferase activities in crude cell extracts from the thermoacidophilic archaeon Thermoplasma acidophilum. We analyzed the modified nucleosides in native initiator and elongator tRNAMet, predicted the candidate genes for the tRNA methyltransferases on the basis of the tRNAMet and tRNALeu sequences, and characterized Trm5, Trm1 and Trm56 by purifying recombinant proteins. We found that the Ta0997, Ta0931, and Ta0836 genes of T. acidophilum encode Trm1, Trm56 and Trm5, respectively. Initiator tRNAMet from T. acidophilum strain HO-62 contained G+, m¹I, and m22G, which were not reported previously in this tRNA, and the m²G26 and m22G26 were formed by Trm1. In the case of elongator tRNAMet, our analysis showed that the previously unidentified G modification at position 26 was a mixture of m²G and m22G, and that they were also generated by Trm1. Furthermore, purified Trm1 and Trm56 could methylate the precursor of elongator tRNAMet, which has an intron at the canonical position. However, the speed of methyl-transfer by Trm56 to the precursor RNA was considerably slower than that to the mature transcript, which suggests that Trm56 acts mainly on the transcript after the intron has been removed. Moreover, cellular arrangements of the tRNA methyltransferases in T. acidophilum are discussed.
- Subjects
RNA modification &; restriction; THERMOPLASMA acidophilum; PROTEIN expression; TRANSFER RNA methyltransferase; ARCHAEBACTERIA; RECOMBINANT protein synthesis
- Publication
International Journal of Molecular Sciences, 2015, Vol 16, Issue 1, p91
- ISSN
1661-6596
- Publication type
Article
- DOI
10.3390/ijms16010091