This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrowReprints and Permissions
Right arrow Copyright Information
Right arrow Books from ASM Press
Right arrow MicrobeWorld
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Adrian, P. V.
Right arrow Articles by Black, T. A.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Adrian, P. V.
Right arrow Articles by Black, T. A.

 Previous Article  |  Next Article 

Antimicrobial Agents and Chemotherapy, November 2000, p. 3101-3106, Vol. 44, No. 11
0066-4804/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Evernimicin (SCH27899) Inhibits a Novel Ribosome Target Site: Analysis of 23S Ribosomal DNA Mutants

Peter V. Adrian,1,* Cara Mendrick,2 David Loebenberg,2 Paul McNicholas,2 Karen J. Shaw,2 Keith P. Klugman,1 Roberta S. Hare,2 and Todd A. Black2

Pneumococcal Diseases Research Unit, South African Institute for Medical Research, University of the Witwatersrand, and the Medical Research Council, Johannesburg, South Africa,1 and Schering Plough Research Institute, Kenilworth, New Jersey2

Received 5 June 2000/Returned for modification 25 July 2000/Accepted 21 August 2000

Spontaneous mutants of susceptible clinical and laboratory isolates of Streptococcus pneumoniae exhibiting reduced susceptibility to evernimicin (SCH27899; MIC, 0.5 to 4.0 mg/liter) were selected on plates containing evernimicin. Four isolates that did not harbor mutations in rplP (which encodes ribosomal protein L16) were further analyzed. Whole chromosomal DNA or PCR products of the 23S ribosomal DNA (rDNA) operons from these mutants could be used to transform the susceptible S. pneumoniae strain R6 to resistance at frequencies of 10-5 and 10-4, respectively, rates 10- to 100-fold lower than that for a single-allele chromosomal marker. The transformants appeared slowly (48 to 72 h) on selective medium, and primary transformants passaged on nonselective medium produced single colonies that displayed heterogeneous susceptibilities to evernimicin. A single passage on selective medium of colonies derived from a single primary transformant homogenized the resistance phenotype. Sequence analysis of the 23S rDNA and rRNA from the resistant mutants revealed single, unique mutations in each isolate at the equivalent Escherichia coli positions 2469 (A right-arrow C), 2480 (C right-arrow T), 2535 (G right-arrow A), and 2536 (G right-arrow C). The mutations map within two different stems of the peptidyltransferase region of domain V. Because multiple copies of rDNA are present in the chromosome, gene conversion between mutant and wild-type 23S rDNA alleles may be necessary for stable resistance. Additionally, none of the characterized mutants showed cross-resistance to any of a spectrum of protein synthesis inhibitors, suggesting that the target site of evernimicin may be unique.

* Corresponding author. Present address: Laboratory of Pediatrics, Erasmus University Rotterdam, Posbus 1738, 3000 DR Rotterdam, The Netherlands. Phone: 31-10-4087951. Fax: 31-10-4089486. E-mail: adrian{at}kgk.fgg.eur.nl.

Antimicrobial Agents and Chemotherapy, November 2000, p. 3101-3106, Vol. 44, No. 11
0066-4804/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


This article has been cited by other articles:

  • Boumghar-Bourtchai, L., Dhalluin, A., Malbruny, B., Galopin, S., Leclercq, R. (2009). Influence of Recombination on Development of Mutational Resistance to Linezolid in Enterococcus faecalis JH2-2. Antimicrob. Agents Chemother. 53: 4007-4009 [Abstract] [Full Text]  
  • Yassin, A., Mankin, A. S. (2007). Potential New Antibiotic Sites in the Ribosome Revealed by Deleterious Mutations in RNA of the Large Ribosomal Subunit. J. Biol. Chem. 282: 24329-24342 [Abstract] [Full Text]  
  • Wolter, N., Smith, A. M., Farrell, D. J., Klugman, K. P. (2006). Heterogeneous Macrolide Resistance and Gene Conversion in the Pneumococcus. Antimicrob. Agents Chemother. 50: 359-361 [Abstract] [Full Text]  
  • Yassin, A., Fredrick, K., Mankin, A. S. (2005). Deleterious mutations in small subunit ribosomal RNA identify functional sites and potential targets for antibiotics. Proc. Natl. Acad. Sci. USA 102: 16620-16625 [Abstract] [Full Text]  
  • Petsaris, O., Miszczak, F., Gicquel-Bruneau, M., Perrin-Guyomard, A., Humbert, F., Sanders, P., Leclercq, R. (2005). Combined Antimicrobial Resistance in Enterococcus faecium Isolated from Chickens. Appl. Environ. Microbiol. 71: 2796-2799 [Abstract] [Full Text]  
  • Lobritz, M., Hutton-Thomas, R., Marshall, S., Rice, L. B. (2003). Recombination Proficiency Influences Frequency and Locus of Mutational Resistance to Linezolid in Enterococcus faecalis. Antimicrob. Agents Chemother. 47: 3318-3320 [Abstract] [Full Text]  
  • Butaye, P., Devriese, L. A., Haesebrouck, F. (2003). Antimicrobial Growth Promoters Used in Animal Feed: Effects of Less Well Known Antibiotics on Gram-Positive Bacteria. Clin. Microbiol. Rev. 16: 175-188 [Abstract] [Full Text]  
  • Kofoed, C. B., Vester, B. (2002). Interaction of Avilamycin with Ribosomes and Resistance Caused by Mutations in 23S rRNA. Antimicrob. Agents Chemother. 46: 3339-3342 [Abstract] [Full Text]  
  • Zarazaga, M., Tenorio, C., Del Campo, R., Ruiz-Larrea, F., Torres, C. (2002). Mutations in Ribosomal Protein L16 and in 23S rRNA in Enterococcus Strains for Which Evernimicin MICs Differ. Antimicrob. Agents Chemother. 46: 3657-3659 [Abstract] [Full Text]  
  • Belova, L., Tenson, T., Xiong, L., McNicholas, P. M., Mankin, A. S. (2001). A novel site of antibiotic action in the ribosome: Interaction of evernimicin with the large ribosomal subunit. Proc. Natl. Acad. Sci. USA 10.1073/pnas.071527498v1 [Abstract] [Full Text]  
  • McNicholas, P. M., Mann, P. A., Najarian, D. J., Miesel, L., Hare, R. S., Black, T. A. (2001). Effects of Mutations in Ribosomal Protein L16 on Susceptibility and Accumulation of Evernimicin. Antimicrob. Agents Chemother. 45: 79-83 [Abstract] [Full Text]  
  • Belova, L., Tenson, T., Xiong, L., McNicholas, P. M., Mankin, A. S. (2001). A novel site of antibiotic action in the ribosome: Interaction of evernimicin with the large ribosomal subunit. Proc. Natl. Acad. Sci. USA 98: 3726-3731 [Abstract] [Full Text]  


Copyright © 2009 by the American Society for Microbiology.   For an alternate route to Journals.ASM.org, visit: http://intl-journals.asm.org | More Info»