In this study, we identified the Saccharomyces cerevisiae RKM2 gene encoding a methyltransferase that catalyzes the trimethylation of a lysine residue in the N terminus of the large subunit ribosomal protein Rpl12ab. Based on the previous identification of dimethyllysine and trimethyllysine in this protein (Lhoest, J., Lobet, Y., Costers, E., and Colson, C. (1984) Eur. J. Biochem. 141, 585-590) as well as our mass spectrometry and amino acid analyses, we suggested that Rpl12ab was modified by dimethylation at Lys-3 catalyzed by an as yet unidentified enzyme and by trimethylation at Lys-10 by Rkm2. Recently, two published articles have suggested that the Arabidopsis thaliana (Carroll, A. J., Heazlewood, J. L., Ito, J., and Millar, A. H. (2008) Mol. Cell. Proteomics 7, 347-369) and Schizosaccharomyces pombe (Sadaie, M., Shinmyozu, K., and Nakayama, J. (2008) J. Biol. Chem. 283, 7185-7195) homologs of S. cerevisiae Rpl12ab are modified by the dimethylation of the N-terminal proline residue and by the trimethylation of Lys-3. The authors of the Arabidopsis study pointed out that the mass spectral data presented in our work are consistent with the S. cerevisiae protein having the same pattern of methylation as the A. thaliana protein. We have now re-evaluated our mass spectral data and agree that our data are consistent with the dimethylation of Pro-1 and the Rkm2-dependent trimethylation of Lys-3, although the evidence presented in our study does support at least a minor degree of alternative trimethylation at Lys-10 (b8, b9, and y9 ions in the wild-type mass spectrum in Fig. 4). Additional experiments performed in our laboratories by Kristofor Webb (to be published) have confirmed that the major trimethylation site is at Lys-3 rather than at Lys-10. These results suggest that the Rpl12ab proteins in S. cerevisiae, S. pombe, and A. thaliana all share the same modifications at the N terminus: a dimethylproline residue at position 1 and a trimethyllysine residue at position 3. A corrected title is shown above.
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| Evidence ID | Analyze ID | Gene/Complex | Systematic Name/Complex Accession | Qualifier | Gene Ontology Term ID | Gene Ontology Term | Aspect | Annotation Extension | Evidence | Method | Source | Assigned On | Reference |
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| Evidence ID | Analyze ID | Gene | Gene Systematic Name | Phenotype | Experiment Type | Experiment Type Category | Mutant Information | Strain Background | Chemical | Details | Reference |
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| Evidence ID | Analyze ID | Gene | Gene Systematic Name | Disease Ontology Term | Disease Ontology Term ID | Qualifier | Evidence | Method | Source | Assigned On | Reference |
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| Evidence ID | Analyze ID | Regulator | Regulator Systematic Name | Target | Target Systematic Name | Direction | Regulation of | Happens During | Regulator Type | Direction | Regulation Of | Happens During | Method | Evidence | Strain Background | Reference |
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| Site | Modification | Modifier | Source | Reference |
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| Evidence ID | Analyze ID | Interactor | Interactor Systematic Name | Interactor | Interactor Systematic Name | Allele | Assay | Annotation | Action | Phenotype | SGA score | P-value | Source | Reference | Note |
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| Evidence ID | Analyze ID | Interactor | Interactor Systematic Name | Interactor | Interactor Systematic Name | Assay | Annotation | Action | Modification | Source | Reference | Note |
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| Complement ID | Locus ID | Gene | Species | Gene ID | Strain background | Direction | Details | Source | Reference |
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| Evidence ID | Analyze ID | Dataset | Description | Keywords | Number of Conditions | Reference |
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