Identification and characterization of tRNA (Gm18) methyltransferase from <i>Thermus thermophilus</i> HB8: domain structure and conserved amino acid sequence motifs

Genes to Cells - Tập 7 Số 3 - Trang 259-272 - 2002
Hiroyuki Hori1, Tsutomu Suzuki2, Kazumasa Sugawara2, Yorinao Inoue3, Takehiko Shibata4, S. Kuramitsu5,6, Shigeyuki Yokoyama7,6,3, Tairo Oshima8, Kimitsuna Watanabe2
1Department of Biochemistry and Molecular Biology, Nippon Medical School, Bunkyo-ku, Tokyo 113-8602, Japan
2Department of Integrated Biosciences, Graduate School of Frontier Sciences, University of Tokyo, 5-1-5 Kashiwanoha, Kashiwa-shi, Chiba, 277-8562, Japan
3RIKEN Harima Institute/SPring-8, Sayou-gun 679-5148, Japan
4Cellular and Molecular Biology Laboratory, RIKEN Institute, Wako 351-0198, Japan
5Faculty of Science, Osaka University, Toyonaka 560-0043, Japan
6RIKEN Genomic Sciences Center, Yokohama 230-0045, Japan
7Graduate School of Science, University of Tokyo, Tokyo 113-0033, Japan
8Tokyo University of Pharmacy and Life Science, Hachioji 192-0392, Japan

Tóm tắt

AbstractBackground: Transfer RNAs from an extreme thermophile, Thermus thermophilus, commonly possess 2′‐O‐methylguanosine at position 18 (Gm18) in the D‐loop. This modification is post‐transcriptionally introduced by tRNA (Gm18) methyltransferase.Results: Partial amino acid sequence data were obtained from purified T. thermophilus tRNA (Gm18) methyltransferase by peptide sequencing and mass spectrometry. The sequence data were used to screen the T. thermophilus genome database currently in progress, resulting in the identification of the corresponding gene. Purified recombinant enzyme showed a strict specificity for methylation at the 2′‐OH of G18 in tRNA. Sequence alignment with other known or putative methyltransferases elucidates that tRNA (Gm18) methyltransferases have specific conserved region as well as three consensus motifs found in RNA ribose 2′‐O‐methyltransferases. The enzyme truncated at its N and C termini by limited tryptic digestion still retained binding activity for S‐adenosyl‐l‐homocysteine, but lost the catalytic activity.Conclusion: This is the first report describing the identification of a methyltransferase gene of the trmH family through the analysis of a purified protein. Further, our results indicate that a restricted region(s) in the terminal amino acid residues of T. thermophilus tRNA (Gm18) methyltransferase are responsible for tRNA recognition and a main part of the enzyme is allocated for a catalytic core.

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Tài liệu tham khảo

10.1021/ac960916h

Auffinger P., 1998, Modification and Editing of RNA, 569

Barbosa E., 1978, mRNA (nucleoside‐2′‐)‐methyltransferase from vaccinia virus. Purification and physical properties, J. Biol. Chem, 253, 7692, 10.1016/S0021-9258(17)34425-3

Björk G.R., 1992, Transfer RNA in Protein Synthesis, 23

10.1128/9781555818333.ch11

10.1146/annurev.bi.56.070187.001403

10.1016/S1097-2765(00)00035-6

10.1074/jbc.M001854200

10.1017/S1355838299981475

10.1021/bi00699a002

10.1016/0300-9084(91)90143-O

10.1002/j.1460-2075.1986.tb04329.x

10.1038/266235a0

10.1016/0300-9084(96)88097-7

10.1093/nar/24.19.3756

10.1016/S0092-8674(00)81101-0

Hori H., 1989, Effects of modification of 4‐thiouridine in E. coli tRNA (fMet) on its methyl acceptor activity by thermostable Gm‐methylases, J. Biochem, 106, 798, 10.1093/oxfordjournals.jbchem.a122933

10.1074/jbc.273.40.25721

10.1006/abbi.1994.1187

10.1021/bi00119a012

10.1016/0300-9084(96)88118-1

10.1016/S0092-8674(00)81308-2

10.1111/j.1432-1033.1973.tb03130.x

10.1073/pnas.92.25.11921

10.1073/pnas.77.4.1922

Kumagai I., 1982, A thermostable tRNA (guanosine‐2′‐)‐methyltransferase from Thermus thermophilus HB27 and the effect of ribose methylation on the conformational stability of tRNA, J. Biol. Chem, 257, 7388, 10.1016/S0021-9258(18)34389-8

10.1038/227680a0

10.1016/0300-9084(96)88098-9

10.1093/nar/22.12.2183

10.1038/288293a0

10.1016/0300-9084(96)88100-4

10.1006/jmbi.1995.0577

Matsumoto T., 1990, Recognition sites of tRNA by a thermostable tRNA (guanosine‐2′‐)‐methyltransferase from Thermus thermophilus HB27, J. Biochem, 107, 331, 10.1093/oxfordjournals.jbchem.a123047

10.1093/oxfordjournals.jbchem.a121983

10.1093/nar/26.1.196

McCloskey J.A., 1999, The RNA modification database: 1999 update, Nucl. Acids Res, 27, 196, 10.1093/nar/27.1.196

Miura K., 1981, The cap structure in eukaryotic messenger RNA as a mark of a strand carrying protein information, Adv. Biophys, 14, 205

10.1006/jmbi.1996.0391

Persson B.C., 1997, The spoU gene of Escherichia coli, the fourth gene of the spoT operon, is essential for tRNA (Gm18) 2′‐O‐methyltransferase activity, Nucl. Acids Res, 25, 3969, 10.1093/nar/25.20.4093

Sambrook J., 1989, Molecular Cloning, 9.31

10.1006/jmbi.1994.0117

10.1126/science.8266080

10.1016/0006-3002(59)90045-9

10.1093/nar/26.1.148

10.1093/nar/21.13.3011

10.1074/jbc.M100432200

10.1016/0300-9084(96)88099-0

Thompson J., 1982, Site of action of a ribosomal RNA methylase conferring resistance to thiostrepton, J. Biol. Chem, 257, 7915, 10.1016/S0021-9258(18)34268-6

10.1073/pnas.93.25.14480

10.1093/emboj/19.3.317

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Genes to Cells

Tập 7 Số 3

259-272

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