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 Garza-Ramos, G.
Right arrow Articles by Mankin, A.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Garza-Ramos, G.
Right arrow Articles by Mankin, A.

 Previous Article  |  Next Article 

Journal of Bacteriology, December 2001, p. 6898-6907, Vol. 183, No. 23
0021-9193/01/$04.00+0   DOI: 10.1128/JB.183.23.6898-6907.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Binding Site of Macrolide Antibiotics on the Ribosome: New Resistance Mutation Identifies a Specific Interaction of Ketolides with rRNA

Georgina Garza-Ramos,1,dagger Liqun Xiong,1 Ping Zhong,2,Dagger and Alexander Mankin1,*

Center for Pharmaceutical Biotechnology, University of Illinois, Chicago, Illinois 60607,1 and Infectious Disease Research, Abbott Laboratories, Abbott Park, Illinois 600642

Received 31 July 2001/Accepted 19 September 2001

Macrolides represent a clinically important class of antibiotics that block protein synthesis by interacting with the large ribosomal subunit. The macrolide binding site is composed primarily of rRNA. However, the mode of interaction of macrolides with rRNA and the exact location of the drug binding site have yet to be described. A new class of macrolide antibiotics, known as ketolides, show improved activity against organisms that have developed resistance to previously used macrolides. The biochemical reasons for increased potency of ketolides remain unknown. Here we describe the first mutation that confers resistance to ketolide antibiotics while leaving cells sensitive to other types of macrolides. A transition of U to C at position 2609 of 23S rRNA rendered E. coli cells resistant to two different types of ketolides, telithromycin and ABT-773, but increased slightly the sensitivity to erythromycin, azithromycin, and a cladinose-containing derivative of telithromycin. Ribosomes isolated from the mutant cells had reduced affinity for ketolides, while their affinity for erythromycin was not diminished. Possible direct interaction of ketolides with position 2609 in 23S rRNA was further confirmed by RNA footprinting. The newly isolated ketolide-resistance mutation, as well as 23S rRNA positions shown previously to be involved in interaction with macrolide antibiotics, have been modeled in the crystallographic structure of the large ribosomal subunit. The location of the macrolide binding site in the nascent peptide exit tunnel at some distance from the peptidyl transferase center agrees with the proposed model of macrolide inhibitory action and explains the dominant nature of macrolide resistance mutations. Spatial separation of the rRNA residues involved in universal contacts with macrolides from those believed to participate in structure-specific interactions with ketolides provides the structural basis for the improved activity of the broader spectrum group of macrolide antibiotics.

* Corresponding author. Mailing address: Center for Pharmaceutical Biotechnology---M/C 870, University of Illinois, 900 S. Ashland Ave., Rm. 3056, Chicago, IL 60607. Phone: (312) 413-1406. Fax: (312) 413-9303. E-mail: shura{at}uic.edu.

dagger Present address: Departamento de Bioquimica, Facultad de Medicina, Universidad Nacional, Autonoma de Mexico, Mexico City, D.F. 04510, Mexico

Dagger Present address: Givaudan Flavors Corporation, Cincinnati, OH 45216.

Journal of Bacteriology, December 2001, p. 6898-6907, Vol. 183, No. 23
0021-9193/01/$04.00+0   DOI: 10.1128/JB.183.23.6898-6907.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


This article has been cited by other articles:

  • Kouvela, E. C., Kalpaxis, D. L., Wilson, D. N., Dinos, G. P. (2009). Distinct Mode of Interaction of a Novel Ketolide Antibiotic That Displays Enhanced Antimicrobial Activity. Antimicrob. Agents Chemother. 53: 1411-1419 [Abstract] [Full Text]  
  • Petropoulos, A. D., Kouvela, E. C., Dinos, G. P., Kalpaxis, D. L. (2008). Stepwise Binding of Tylosin and Erythromycin to Escherichia coli Ribosomes, Characterized by Kinetic and Footprinting Analysis. J. Biol. Chem. 283: 4756-4765 [Abstract] [Full Text]  
  • Cruz-Vera, L. R., New, A., Squires, C., Yanofsky, C. (2007). Ribosomal Features Essential for tna Operon Induction: Tryptophan Binding at the Peptidyl Transferase Center. J. Bacteriol. 189: 3140-3146 [Abstract] [Full Text]  
  • Struble, J. M., Gill, R. T. (2006). Reverse Engineering Antibiotic Sensitivity in a Multidrug-Resistant Pseudomonas aeruginosa Isolate.. Antimicrob. Agents Chemother. 50: 2506-2515 [Abstract] [Full Text]  
  • Hisanaga, T., Hoban, D. J., Zhanel, G. G. (2005). Mechanisms of resistance to telithromycin in Streptococcus pneumoniae. J Antimicrob Chemother 56: 447-450 [Abstract] [Full Text]  
  • Mamelli, L., Prouzet-Mauleon, V., Pages, J.-M., Megraud, F., Bolla, J.-M. (2005). Molecular basis of macrolide resistance in Campylobacter: role of efflux pumps and target mutations. J Antimicrob Chemother 56: 491-497 [Abstract] [Full Text]  
  • Yan, K., Hunt, E., Berge, J., May, E., Copeland, R. A., Gontarek, R. R. (2005). Fluorescence Polarization Method To Characterize Macrolide-Ribosome Interactions. Antimicrob. Agents Chemother. 49: 3367-3372 [Abstract] [Full Text]  
  • Nash, K. A., Zhang, Y., Brown-Elliott, B. A., Wallace, R. J. Jr (2005). Molecular basis of intrinsic macrolide resistance in clinical isolates of Mycobacterium fortuitum. J Antimicrob Chemother 55: 170-177 [Abstract] [Full Text]  
  • Abbanat, D., Webb, G., Foleno, B., Li, Y., Macielag, M., Montenegro, D., Wira, E., Bush, K. (2005). In Vitro Activities of Novel 2-Fluoro-Naphthyridine-Containing Ketolides. Antimicrob. Agents Chemother. 49: 309-315 [Abstract] [Full Text]  
  • Novotny, G. W., Jakobsen, L., Andersen, N. M., Poehlsgaard, J., Douthwaite, S. (2004). Ketolide Antimicrobial Activity Persists after Disruption of Interactions with Domain II of 23S rRNA. Antimicrob. Agents Chemother. 48: 3677-3683 [Abstract] [Full Text]  
  • Ackermann, G., Rodloff, A. C. (2003). Drugs of the 21st century: telithromycin (HMR 3647)--the first ketolide. J Antimicrob Chemother 51: 497-511 [Abstract] [Full Text]  
  • Dinos, G. P., Connell, S. R., Nierhaus, K. H., Kalpaxis, D. L. (2003). Erythromycin, Roxithromycin, and Clarithromycin: Use of Slow-Binding Kinetics to Compare Their in Vitro Interaction with a Bacterial Ribosomal Complex Active in Peptide Bond Formation. Mol. Pharmacol. 63: 617-623 [Abstract] [Full Text]  
  • Pereyre, S., Gonzalez, P., de Barbeyrac, B., Darnige, A., Renaudin, H., Charron, A., Raherison, S., Bebear, C., Bebear, C. M. (2002). Mutations in 23S rRNA Account for Intrinsic Resistance to Macrolides in Mycoplasma hominis and Mycoplasma fermentans and for Acquired Resistance to Macrolides in M. hominis. Antimicrob. Agents Chemother. 46: 3142-3150 [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»