Actin-binding protein ABP140 is a methyltransferase for 3-methylcytidine at position 32 of tRNAs in Saccharomyces cerevisiae.

Akiko Noma, Sanghyun Yi, Takayuki Katoh, Yoshimi Takai, Takeo Suzuki, Tsutomu Suzuki

Department of Chemistry and Biotechnology, Graduate School of Engineering, University of Tokyo, Bunkyo-ku, Tokyo 113-8656, Japan.

RNA (New York, N.Y.) 2011 Jun

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Transfer RNAs contain various modified nucleotides that are introduced enzymatically at the post-transcriptional level. In Saccharomyces cerevisiae, 3-methylcytidine (m³C) is found at position 32 of the tRNAs for Thr and Ser. We used a systematic reverse genetic approach combined with mass spectrometry (ribonucleome analysis), and identified the actin-binding protein ABP140 as the protein responsible for m³C formation in both tRNA(Thr1) and tRNA(Ser1). ABP140 consists of an N-terminal actin-binding sequence and a C-terminal S-adenosylmethionine (Ado-Met) binding motif. Deletion of the actin-binding sequence in ABP140 did not affect m³C formation, indicating that subcellular localization of ABP140 to actin filaments is not involved in tRNA modification. m³C formation in tRNA(Thr1) could be reconstituted using recombinant Abp140p in the presence of Ado-Met, whereas m³C did not form in tRNA(Ser1) in vitro, indicating the absence of a factor(s) required for tRNA(Ser1) m³C formation. Thus, ABP140 has been designated TRM140 according to the preferred nomenclature. In addition, we observed a specific reduction of m³C formation in HeLa cells by siRNA-mediated knock down of the human ortholog of TRM140.

Citation

Akiko Noma, Sanghyun Yi, Takayuki Katoh, Yoshimi Takai, Takeo Suzuki, Tsutomu Suzuki. Actin-binding protein ABP140 is a methyltransferase for 3-methylcytidine at position 32 of tRNAs in Saccharomyces cerevisiae. RNA (New York, N.Y.). 2011 Jun;17(6):1111-9