The hemK gene in Escherichia coli encodes the N(5)-glutamine methyltransferase that modifies peptide release factors.
Class 1 peptide release factors (RFs) in Escherichia coli are N(5)-methylated on the glutamine residue of the universally conserved GGQ motif. One other protein alone has been shown to contain N(5)-methylglutamine: E.coli ribosomal protein L3. We identify the L3 methyltransferase as YfcB and show that it methylates ribosomes from a yfcB strain in vitro, but not RF1 or RF2. HemK, a close orthologue of YfcB, is shown to methylate RF1 and RF2 in vitro. hemK is immediately downstream of and co-expressed with prfA. Its deletion in E.coli K12 leads to very poor growth on rich media and abolishes methylation of RF1. The activity of unmethylated RF2 from K12 strains is extremely low due to the cumulative effects of threonine at position 246, in place of alanine or serine present in all other bacterial RFs, and the lack of N(5)-methylation of Gln252. Fast-growing spontaneous revertants in hemK K12 strains contain the mutations Thr246Ala or Thr246Ser in RF2. HemK and YfcB are the first identified methyltransferases modifying glutamine, and are widely distributed in nature.
Heurgué-Hamard
Valérie
V
UPR9073 du CNRS, Institut de Biologie Physico-Chimique, 13 rue Pierre et Marie Curie, Paris 75005, France.
Champ
Stéphanie
S
Engström
Ake
A
Ehrenberg
Måns
M
Buckingham
Richard H
RH
D000595:Amino Acid Sequence; D004926:Escherichia coli / Q000235:genetics* / Q000254:growth & development; D029968:Escherichia coli Proteins / Q000235:genetics*; D005798:Genes, Bacterial; D008745:Methylation; D008969:Molecular Sequence Data; D010455:Peptides / Q000378:metabolism; D011496:Protein Methyltransferases / Q000235:genetics*; D017386:Sequence Homology, Amino Acid
D029968:Escherichia coli Proteins; D010455:Peptides; D011496:Protein Methyltransferases; C092404:prmC protein, E coli