Home LiteratureArticle Details
PMID: 11847124 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

The hemK gene in Escherichia coli encodes the N(5)-glutamine methyltransferase that modifies peptide release factors.

The EMBO journal ·Vol. 21 ·No. 4 ·2002-02-15 ·Pages 769-78

Heurgué-Hamard V, Champ S, Engström A, Ehrenberg M, Buckingham RH

Abstract

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.

MeSH Terms
Amino Acid Sequence Escherichia coli/genetics,growth & development Escherichia coli Proteins/genetics Genes, Bacterial Methylation Molecular Sequence Data Peptides/metabolism Protein Methyltransferases/genetics Sequence Homology, Amino Acid
Chemicals
Escherichia coli Proteins Peptides Protein Methyltransferases prmC protein, E coli
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Heurgué-Hamard Valérie
UPR9073 du CNRS, Institut de Biologie Physico-Chimique, 13 rue Pierre et Marie Curie, Paris 75005, France.
Champ Stéphanie
Engström Ake
Ehrenberg Måns
Buckingham Richard H
References (42)
42 references, click to expand
  1. The crystal structure of human eukaryotic release factor eRF1--mechanism of stop codon recognition and peptidyl-tRNA hydrolysis.
    Cell. 2000 Feb 4;100(3):311-21 PMID: 10676813
  2. Acetylornithinase of Escherichia coli: partial purification and some properties.
    J Biol Chem. 1956 Jan;218(1):97-106 PMID: 13278318
  3. Is the HemK family of putative S-adenosylmethionine-dependent methyltransferases a "missing" zeta subfamily of adenine methyltransferases? A hypothesis.
    IUBMB Life. 1999 Sep;48(3):247-9 PMID: 10690633
  4. The accuracy of codon recognition by polypeptide release factors.
    Proc Natl Acad Sci U S A. 2000 Feb 29;97(5):2046-51 PMID: 10681447
  5. Translation termination in eukaryotes: polypeptide release factor eRF1 is composed of functionally and structurally distinct domains.
    RNA. 2000 Mar;6(3):381-90 PMID: 10744022
  6. Predicting subcellular localization of proteins based on their N-terminal amino acid sequence.
    J Mol Biol. 2000 Jul 21;300(4):1005-16 PMID: 10891285
  7. [Mutation of a glutamine residue in the universal tripeptide GGQ in human eRF1 termination factor does not cause complete loss of its activity].
    Mol Biol (Mosk). 2000 Sep-Oct;34(5):899-900 PMID: 11033819
  8. Escherichia coli TehB requires S-adenosylmethionine as a cofactor to mediate tellurite resistance.
    J Bacteriol. 2000 Nov;182(22):6509-13 PMID: 11053398
  9. Translational termination comes of age.
    Trends Biochem Sci. 2000 Nov;25(11):561-6 PMID: 11084369
  10. DNA cloning by homologous recombination in Escherichia coli.
    Nat Biotechnol. 2000 Dec;18(12):1314-7 PMID: 11101815
  11. A post-translational modification in the GGQ motif of RF2 from Escherichia coli stimulates termination of translation.
    EMBO J. 2000 Dec 15;19(24):6900-7 PMID: 11118225
  12. AdoMet-dependent methylation, DNA methyltransferases and base flipping.
    Nucleic Acids Res. 2001 Sep 15;29(18):3784-95 PMID: 11557810
  13. Methylation of the ribosomal proteins in Escherichia coli. Nature and stoichiometry of the methylated amino acids in 50S ribosomal proteins.
    Biochemistry. 1975 Feb 11;14(3):468-77 PMID: 1089427
  14. Genetics of ribosomal protein methylation in Escherichia coli. II. A mutant lacking a new type of methylated amino acid, N5-methylglutamine, in protein L3.
    Mol Gen Genet. 1977 Jul 20;154(2):175-80 PMID: 331083
  15. Genetic studies of the lac repressor. III. Additional correlation of mutational sites with specific amino acid residues.
    J Mol Biol. 1977 Dec 15;117(3):525-67 PMID: 609095
  16. The primary structure of ribosomal protein L3 from Escherichia coli 70 S ribosomes.
    FEBS Lett. 1978 Dec 15;96(2):301-5 PMID: 365579
  17. Synthesis and degradation of termination and premature-termination fragments of beta-galactosidase in vitro and in vivo.
    J Mol Biol. 1978 Nov 15;125(4):407-32 PMID: 105143
  18. Genetics of ribosomal protein methylation in Escherichia coli. III. Map position of two genes, prmA and prmB, governing methylation of proteins L11 and L3.
    Mol Gen Genet. 1979 Feb 1;169(3):245-50 PMID: 372746
  19. Use of bacteriophage T7 RNA polymerase to direct selective high-level expression of cloned genes.
    J Mol Biol. 1986 May 5;189(1):113-30 PMID: 3537305
  20. Tightly regulated tac promoter vectors useful for the expression of unfused and fused proteins in Escherichia coli.
    Gene. 1988 Sep 30;69(2):301-15 PMID: 3069586
  21. Construction of versatile low-copy-number vectors for cloning, sequencing and gene expression in Escherichia coli.
    Gene. 1991 Apr;100:195-9 PMID: 2055470
  22. PATMAT: a searching and extraction program for sequence, pattern and block queries and databases.
    Comput Appl Biosci. 1992 Jun;8(3):249-54 PMID: 1633566
  23. Release factor-dependent false stops are infrequent in Escherichia coli.
    J Mol Biol. 1993 Mar 5;230(1):41-50 PMID: 8450549
  24. A mutational analysis of the two motifs common to adenine methyltransferases.
    EMBO J. 1994 Aug 15;13(16):3902-8 PMID: 8070417
  25. CLUSTAL W: improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position-specific gap penalties and weight matrix choice.
    Nucleic Acids Res. 1994 Nov 11;22(22):4673-80 PMID: 7984417
  26. Cloning and sequencing of a previously unidentified gene that is involved in the biosynthesis of heme in Escherichia coli.
    Gene. 1995 Feb 3;153(1):67-70 PMID: 7883187
  27. Ribosomal protein methylation in Escherichia coli: the gene prmA, encoding the ribosomal protein L11 methyltransferase, is dispensable.
    Mol Microbiol. 1994 Dec;14(5):947-58 PMID: 7715456
  28. Gene disruption in Escherichia coli: TcR and KmR cassettes with the option of Flp-catalyzed excision of the antibiotic-resistance determinant.
    Gene. 1995 May 26;158(1):9-14 PMID: 7789817
  29. Structure-based sequence alignment of three AdoMet-dependent DNA methyltransferases.
    Gene. 1995 May 19;157(1-2):135-8 PMID: 7607477
  30. Fitting a mixture model by expectation maximization to discover motifs in biopolymers.
    Proc Int Conf Intell Syst Mol Biol. 1994;2:28-36 PMID: 7584402
  31. Automated construction and graphical presentation of protein blocks from unaligned sequences.
    Gene. 1995 Oct 3;163(2):GC17-26 PMID: 7590261
  32. Structure-guided analysis reveals nine sequence motifs conserved among DNA amino-methyltransferases, and suggests a catalytic mechanism for these enzymes.
    J Mol Biol. 1995 Nov 3;253(4):618-32 PMID: 7473738
  33. Sequencing and analysis of bacterial genomes.
    Curr Biol. 1996 Apr 1;6(4):404-16 PMID: 8723345
  34. Gene order is not conserved in bacterial evolution.
    Trends Genet. 1996 Aug;12(8):289-90 PMID: 8783936
  35. Identification and analysis of genes from Streptomyces pristinaespiralis encoding enzymes involved in the biosynthesis of the 4-dimethylamino-L-phenylalanine precursor of pristinamycin I.
    Mol Microbiol. 1997 Jan;23(2):191-202 PMID: 9044253
  36. Functional specificity of amino acid at position 246 in the tRNA mimicry domain of bacterial release factor 2.
    Biochimie. 1996;78(11-12):935-43 PMID: 9150870
  37. Functional roles of conserved amino acid residues in DNA methyltransferases investigated by site-directed mutagenesis of the EcoRV adenine-N6-methyltransferase.
    J Biol Chem. 1998 Jul 10;273(28):17333-42 PMID: 9651316
  38. A direct estimation of the context effect on the efficiency of termination.
    J Mol Biol. 1998 Dec 4;284(3):579-90 PMID: 9826500
  39. Functional analysis of the hemK gene product involvement in protoporphyrinogen oxidase activity in yeast.
    FEMS Microbiol Lett. 1999 Apr 1;173(1):175-82 PMID: 10220893
  40. Mutations in the highly conserved GGQ motif of class 1 polypeptide release factors abolish ability of human eRF1 to trigger peptidyl-tRNA hydrolysis.
    RNA. 1999 Aug;5(8):1014-20 PMID: 10445876
  41. Shutdown in protein synthesis due to the expression of mini-genes in bacteria.
    J Mol Biol. 1999 Aug 27;291(4):745-59 PMID: 10452886
  42. A tripeptide 'anticodon' deciphers stop codons in messenger RNA.
    Nature. 2000 Feb 10;403(6770):680-4 PMID: 10688208
Article Info
Journal
The EMBO journal
Abbr.
EMBO J
ISSN
0261-4189
DOI
10.1093/emboj/21.4.769
Published
2002-02-15
Pages
769-78
Language
English
Region
England
NLM ID
8208664
PMCID
PMC125846
Subset
IM
External Links
PubMed source DOI: 10.1093/emboj/21.4.769 PMC Full Text View journal details
Analysis Services
Analysis Services

Contact

No. 2 Wenbo Road, Zhangqiu District, Jinan, Shandong

Qilu Normal University · Genelibs Bioinformatics Lab

750 Shunhua Rd, Jinan

2F, Bldg F, University Science Park

Tel: 0531-88819269

WeChat Official Account

Follow our WeChat subscription account for real-time updates and the latest in medical and biological research.


Business Email

E-mail: [email protected]

Copyright © 2026.Genelibs Jinan Honghui Biotechnology Co., Ltd. 鲁ICP备20007159号-2