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PMID:22522802

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Citation

Couttas, TA, Raftery, MJ, Padula, MP, Herbert, BR and Wilkins, MR (2012) Methylation of translation-associated proteins in Saccharomyces cerevisiae: Identification of methylated lysines and their methyltransferases. Proteomics 12:960-72

Abstract

This study aimed to identify sites of lysine methylation in Saccharomyces cerevisiae and the associated methyltransferases. Hexapeptide ligand affinity chromatography was used to normalize the abundance levels of proteins in whole cell lysate. MS/MS, in association with antibody-based detection, was then used to identify lysine methylated proteins and the precise sites of modification. Lysine methylation was found on the proteins elongation factor (EF) 1-α, 2, and 3A, as well as ribosomal proteins 40S S18-A/B, 60S L11-A/B, L18-A/B, and L42-A/B. Precise sites were mapped in all cases. Single-gene knockouts of known and putative methyltransferase(s), in association with MS/MS, showed that EF1-α is monomethylated by Efm1 at lysin 30 and dimethylated by See1 at lysine 316. Methyltransferase Rkm1 was found to monomethylate 40S ribosomal protein S18-A/B at lysine 48. Knockout analysis also revealed that putative methyltransferase YBR271W affects the methylation of proteins EF2 and 3A; this was detected by Western blotting and immunodetection. This methyltransferase shows strong interspecies conservation and a tryptophan-containing motif associated with its active site. We suggest that enzyme YBR271W is named EF methyltransferase 2 (Efm2), in line with the recent naming of YHL039W as Efm1.

Links

PubMed Online version:10.1002/pmic.201100570

Keywords


Significance

Annotations

Gene product Qualifier GO Term Evidence Code with/from Aspect Extension Notes Status

YEAST:EFM2

enables

GO:0016279: protein-lysine N-methyltransferase activity

ECO:0000315: mutant phenotype evidence used in manual assertion

F

Seeded From UniProt

complete

YEAST:EFM1

enables

GO:0016279: protein-lysine N-methyltransferase activity

ECO:0000315: mutant phenotype evidence used in manual assertion

F

  • has_input:(UniProtKB:P02994)

Seeded From UniProt

complete

YEAST:EFM1

involved_in

GO:0018026: peptidyl-lysine monomethylation

ECO:0000315: mutant phenotype evidence used in manual assertion

P

  • has_input:(UniProtKB:P02994)

Seeded From UniProt

complete

YEAST:RKM1

involved_in

GO:0018026: peptidyl-lysine monomethylation

ECO:0000315: mutant phenotype evidence used in manual assertion

P

  • has_input:(UniProtKB:P0CX55)|has_input:(UniProtKB:P0CX56)

Seeded From UniProt

complete

YEAST:SEE1

involved_in

GO:0018027: peptidyl-lysine dimethylation

ECO:0000315: mutant phenotype evidence used in manual assertion

P

has_direct_input:(UniProtKB:P02994)

Seeded From UniProt

complete


See also

References

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