1887

Abstract

Peptide deformylation is an essential process in eubacteria. The peptide deformylase Def has been suggested to be an attractive target for antibacterial drug discovery. Some eubacteria including medically important pathogens possess two like genes. Until now, the functionality of both genes has been tested only in with the result that one gene copy was functional. Here, expression of two functional -like gene products in is demonstrated. Besides the gene, which is chromosomally located close to the formyltransferase gene and which was overexpressed and biochemically tested previously, possesses a second -like gene, called . The encoded protein is 32% identical to the gene product. It was shown that either or had to be present for growth of in rich medium (each was individually dispensable). Studies with a / double deletion strain with xylose-inducible copy demonstrated that, besides , the gene is a second cellular target of deformylase inhibitors such as the antibiotic actinonin. The gene products exhibited similar enzymic properties, exemplified by similar inhibition efficacy of actinonin in biochemical assays. Antibiotic susceptibility tests with different deletion strains and Northern analyses indicated that YkrB is probably the predominant deformylase in . It was shown that duplication of the deformylase function does not lead to an increased actinonin-resistance frequency in comparison to mutants carrying only one deformylase gene.

Loading

Article metrics loading...

/content/journal/micro/10.1099/00221287-147-7-1783
2001-07-01
2024-03-29
Loading full text...

Full text loading...

/deliver/fulltext/micro/147/7/1471783a.html?itemId=/content/journal/micro/10.1099/00221287-147-7-1783&mimeType=html&fmt=ahah

References

  1. Adams J. M. 1968; On the release of the formyl group from nascent protein. J Mol Biol 33:571–589 [CrossRef]
    [Google Scholar]
  2. Anagnostopoulos C., Spizizen J. 1961; Requirements for transformation in Bacillus subtilis . J Bacteriol 81:741–746
    [Google Scholar]
  3. Belouski E., Gui L., Rudolph F. B., Bennett G. N. 1998; Complementation of an Escherichia coli polypeptide deformylase mutant with a gene from Clostridium acetobutylicum ATCC 824. Curr Microbiol 36:248–249 [CrossRef]
    [Google Scholar]
  4. Bradford M. M. 1976; A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254 [CrossRef]
    [Google Scholar]
  5. Braun H. P., Schmitz U. K. 1993; Purification and sequencing of cytochrome b from potato reveals methionine cleavage of a mitochondrially encoded protein. FEBS Lett 316:128–132 [CrossRef]
    [Google Scholar]
  6. Chen D. Z., Patel D. V., Hackbarth C. J. & 9 other authors; 2000; Actinonin, a naturally occurring antibacterial agent, is a potent deformylase inhibitor. Biochemistry 39:1256–1262 [CrossRef]
    [Google Scholar]
  7. Dardel F., Ragusa S., Lazennec C., Blanquet S., Meinnel T. 1998; Solution structure of nickel-peptide deformylase. J Mol Biol 280:501–513 [CrossRef]
    [Google Scholar]
  8. Durand D. J., Gordon Green B., O’Connell J. F., Grant S. K. 1999; Peptide aldehyde inhibitors of bacterial peptide deformylases. Arch Biochem Biophys 367:297–302 [CrossRef]
    [Google Scholar]
  9. Evans V. J., Liyanage H., Ravagnani A., Young M., Kashket E. R. 1998; Truncation of peptide deformylase reduces the growth rate and stabilizes solvent production in Clostridium beijerinckii NCIMB 8052. Appl Environ Microbiol 64:1780–1785
    [Google Scholar]
  10. Ferrari F. A., Nguyen A., Lang D., Hoch J. A. 1983; Construction and properties of an integrable plasmid for Bacillus subtilis . J Bacteriol 154:1513–1515
    [Google Scholar]
  11. Giglione C., Pierre M., Meinnel T. 2000; Peptide deformylase as a target for new generation, broad spectrum antimicrobial agents. Mol Microbiol 36:1197–1205
    [Google Scholar]
  12. Guerout-Fleury A. M., Frandsen N., Stragier P. 1996; Plasmids for ectopic integration in Bacillus subtilis . Gene 180:57–61 [CrossRef]
    [Google Scholar]
  13. Harwood C. R., Cutting S. M. 1990 Molecular Biological Methods for Bacillus Chichester: Wiley;
    [Google Scholar]
  14. Huntington K. M., Yi T., Wei Y., Pei D. 2000; Synthesis and antibacterial activity of peptide deformylase inhibitors. Biochemistry 39:4543–4551 [CrossRef]
    [Google Scholar]
  15. Itaya M., Kondo K., Tanaka T. 1989; A neomycin resistance gene cassette selectable in a single copy state in the Bacillus subtilis chromosome. Nucleic Acids Res 17:4410 [CrossRef]
    [Google Scholar]
  16. Kim L., Mogk A., Schumann W. 1996; A xylose-inducible Bacillus subtilis integration vector and its application. Gene 181:71–76 [CrossRef]
    [Google Scholar]
  17. Kozak M. 1983; Comparison of initiation of protein synthesis in procaryotes, eucaryotes, and organelles. Microbiol Rev 47:1–45
    [Google Scholar]
  18. Leiting B., Marsilio F., O’Connell J. F. 1998; Predictable deuteration of recombinant proteins expressed in Escherichia coli . Anal Biochem 265:351–355 [CrossRef]
    [Google Scholar]
  19. Margolis P. S., Hackbarth C. J., Young D. C., Wang W., Chen D., Yuan Z., White R., Trias J. 2000; Peptide deformylase in Staphylococcus aureus : resistance to inhibition is mediated by mutations in the formyltransferase gene. Antimicrob Agents Chemother 44:1825–1831 [CrossRef]
    [Google Scholar]
  20. Mazel D., Pochet S., Marliere P. 1994; Genetic characterization of polypeptide deformylase, a distinctive enzyme of eubacterial translation. EMBO J 13:914–923
    [Google Scholar]
  21. Mazel D., Coic E., Blanchard S., Saurin W., Marliere P. 1997; A survey of polypeptide deformylase function throughout the eubacterial lineage. J Mol Biol 266:939–949 [CrossRef]
    [Google Scholar]
  22. Meinnel T. 2000; Peptide deformylase of eukaryotic protists: a target for new antiparasitic agents?. Parasitol Today 16:165–168 [CrossRef]
    [Google Scholar]
  23. Meinnel T., Blanquet S. 1993; Evidence that peptide deformylase and methionyl-tRNA(fMet) formyltransferase are encoded within the same operon in Escherichia coli . J Bacteriol 175:7737–7740
    [Google Scholar]
  24. Meinnel T., Blanquet S. 1994; Characterization of the Thermus thermophilus locus encoding peptide deformylase and methionyl-tRNA(fMet) formyltransferase. J Bacteriol 176:7387–7390
    [Google Scholar]
  25. Meinnel T., Lazennec C., Villoing S., Blanquet S. 1997; Structure-function relationships within the peptide deformylase family. Evidence for a conserved architecture of the active site involving three conserved motifs and a metal ion. J Mol Biol 267:749–761 [CrossRef]
    [Google Scholar]
  26. Rygus T., Hillen W. 1992; Catabolite repression of the xyl operon in Bacillus megaterium . J Bacteriol 174:3049–3055
    [Google Scholar]
  27. Sambrook J., Fritsch E. F., Maniatis T. 1989 Molecular Cloning: a Laboratory Manual , 2nd edn. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory;
    [Google Scholar]
  28. Shanklin J., DeWitt N. D., Flanagan J. M. 1995; The stroma of higher plant plastids contain ClpP and ClpC, functional homologs of Escherichia coli ClpP and ClpA: an archetypal two-component ATP-dependent protease. Plant Cell 7:1713–1722
    [Google Scholar]
  29. Sherman F., Stewart J. W., Tsunasawa S. 1985; Methionine or not methionine at the beginning of a protein. Bioessays 3:27–31 [CrossRef]
    [Google Scholar]
  30. Solbiati J., Chapman-Smith A., Miller J. L., Miller C. G., Cronan J. E. Jr 1999; Processing of the N termini of nascent polypeptide chains requires deformylation prior to methionine removal. J Mol Biol 290:607–614 [CrossRef]
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/00221287-147-7-1783
Loading
/content/journal/micro/10.1099/00221287-147-7-1783
Loading

Data & Media loading...

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error