Published online 21 March 2005
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Modification at position 9 with 1-methyladenosine is crucial for structure and function of nematode mitochondrial tRNAs lacking the entire T-arm
1Department of Integrated Biosciences, Graduate School of Frontier Sciences, The University of Tokyo Kashiwa, Chiba 277-8562, Japan 2Department of Chemistry and Biotechnology, Graduate School of Engineering, The University of Tokyo Bunkyo-ku, Tokyo 113-8656, Japan
*To whom correspondence should be addressed at Biological Information Research Center, National Institute of Advanced Industrial Science and Technology, 2-41-6 Aomi, Koto-ku, Tokyo 135-0064, Japan. Tel: +81 3 3599 8106; Fax: +81 3 5530 2064; Email: kwatanab{at}jbirc.aist.go.jp
Received February 2, 2005. Revised February 28, 2005. Accepted February 28, 2005.
The mitochondria of the nematode Ascaris suum have tRNAs with unusual secondary structures that lack either the T-arm or D-arm found in most other organisms. Of the twenty-two tRNA species present in the mitochondria of A.suum, twenty lack the entire T-arm and two serine tRNAs lack the D-arm. To understand how such unusual tRNAs work in the nematode mitochondrial translation system, we analyzed post-transcriptional modifications of 11 mitochondrial tRNA species purified from A.suum, 10 of which lacked a T-arm and one of which lacked a D-arm. The most characteristic feature of nematode mitochondrial tRNAs lacking a T-arm was the presence of 1-methyladenosine at position 9 (m1A9). Synthesis of T-armless tRNAs with or without the modified nucleoside showed that T-armless tRNAs without the modification had much lower aminoacylation and EF-Tu-binding activities than native tRNAs. The addition of a single methyl group to A9 of these tRNAs was sufficient to restore nearly native levels of aminoacylation and EF-Tu-binding activity as well as tertiary structure, suggesting that m1A9 is a key residue for the activity of T-armless tRNAs. Thus, m1A9 is indispensable for the structure and function of T-armless tRNAs of nematode mitochondrial origin.
Correspondence may addressed to Takashi Ohtsuki, Department of Bioscience and Biotechnology, Okayama University, 3-1-1 Tsushimanaka, Okayama 700-8530, Japan. Tel: +81 86 251 8220; Fax: +81 86 251 8219; Email: ohtsuk{at}cc.okayama-u.ac.jp
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