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 Olesky, M.
Right arrow Articles by Nicholas, R. A.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Olesky, M.
Right arrow Articles by Nicholas, R. A.

 Previous Article  |  Next Article 

Antimicrobial Agents and Chemotherapy, September 2002, p. 2811-2820, Vol. 46, No. 9
0066-4804/02/$04.00+0     DOI: 10.1128/AAC.46.9.2811-2820.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Identification and Analysis of Amino Acid Mutations in Porin IB That Mediate Intermediate-Level Resistance to Penicillin and Tetracycline in Neisseria gonorrhoeae

Melanie Olesky,1 Marcia Hobbs,2,3 and Robert A. Nicholas1*

Departments of Pharmacology,1 Medicine,2 Microbiology and Immunology, The University of North Carolina at Chapel Hill, Chapel Hill, North Carolina 27599-73653

Received 14 March 2002/ Returned for modification 15 May 2002/ Accepted 11 June 2002

PenB is the third resistance determinant in the stepwise acquisition of multiple resistance genes in chromosomally mediated resistant Neisseria gonorrhoeae (CMRNG). Alterations in porIB, one of two alleles at the por locus that encodes the outer membrane protein porin IB (PIB), were recently reported to be responsible for the increased resistance to penicillin and tetracycline conferred by penB, but the specific mutations conferring antibiotic resistance were not identified experimentally. To determine which amino acids in PIB confer increased resistance, we transformed a recipient strain with chimeras of the porIB genes from strains FA1090 and FA140 (penB2). These studies revealed that two amino acid changes, G120D and A121D, were both necessary and sufficient to confer increased resistance to penicillin and tetracycline. Site-saturation and site-directed mutagenesis of Gly-120 and Ala-121 revealed that both a single mutation, G120K, and the double mutations G120R A121H and G120P A121P also conferred antibiotic resistance to the recipient strain. The identical mutations in PIA increased penicillin and tetracycline resistance either moderately or not at all. Analysis of porIB genes present in the GenBank database from 51 clinical isolates demonstrated that lysine and aspartate mutations at positions 120 and/or 121 also occur in nature. These studies demonstrate that charged amino acids at positions 120 and 121 in PIB are highly preferential for conferring resistance to penicillin and tetracycline in N. gonorrhoeae.

* Corresponding author. Mailing address: Department of Pharmacology, The University of North Carolina at Chapel Hill, Chapel Hill, NC 27599-7364. Phone: (919) 966-6547. Fax: (919) 966-5640. E-mail: nicholas{at}med.unc.edu.

Antimicrobial Agents and Chemotherapy, September 2002, p. 2811-2820, Vol. 46, No. 9
0066-4804/02/$04.00+0     DOI: 10.1128/AAC.46.9.2811-2820.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Kugelman, G., Tapsall, J. W., Goire, N., Syrmis, M. W., Limnios, A., Lambert, S. B., Nissen, M. D., Sloots, T. P., Whiley, D. M. (2009). Simple, Rapid, and Inexpensive Detection of Neisseria gonorrhoeae Resistance Mechanisms Using Heat-Denatured Isolates and SYBR Green-Based Real-Time PCR. Antimicrob. Agents Chemother. 53: 4211-4216 [Abstract] [Full Text]  
  • Zhao, S., Duncan, M., Tomberg, J., Davies, C., Unemo, M., Nicholas, R. A. (2009). Genetics of Chromosomally Mediated Intermediate Resistance to Ceftriaxone and Cefixime in Neisseria gonorrhoeae. Antimicrob. Agents Chemother. 53: 3744-3751 [Abstract] [Full Text]  
  • Powell, A. J., Tomberg, J., Deacon, A. M., Nicholas, R. A., Davies, C. (2009). Crystal Structures of Penicillin-binding Protein 2 from Penicillin-susceptible and -resistant Strains of Neisseria gonorrhoeae Reveal an Unexpectedly Subtle Mechanism for Antibiotic Resistance. J. Biol. Chem. 284: 1202-1212 [Abstract] [Full Text]  
  • Vernel-Pauillac, F., Nandi, S., Nicholas, R. A., Goarant, C. (2008). Genotyping as a Tool for Antibiotic Resistance Surveillance of Neisseria gonorrhoeae in New Caledonia: Evidence of a Novel Genotype Associated with Reduced Penicillin Susceptibility. Antimicrob. Agents Chemother. 52: 3293-3300 [Abstract] [Full Text]  
  • Liao, M., Bell, K., Gu, W.-M., Yang, Y., Eng, N. F., Fu, W., Wu, L., Zhang, C.-G., Chen, Y., Jolly, A. M., Dillon, J.-A. R. (2008). Clusters of circulating Neisseria gonorrhoeae strains and association with antimicrobial resistance in Shanghai. J Antimicrob Chemother 61: 478-487 [Abstract] [Full Text]  
  • Lindberg, R., Fredlund, H., Nicholas, R., Unemo, M. (2007). Neisseria gonorrhoeae Isolates with Reduced Susceptibility to Cefixime and Ceftriaxone: Association with Genetic Polymorphisms in penA, mtrR, porB1b, and ponA. Antimicrob. Agents Chemother. 51: 2117-2122 [Abstract] [Full Text]  
  • Vernel-Pauillac, F., Merien, F. (2006). A Novel Real-Time Duplex PCR Assay for Detecting penA and ponA Genotypes in Neisseria gonorrhoeae: Comparison with Phenotypes Determined by the E-Test. Clin. Chem. 52: 2294-2296 [Abstract] [Full Text]  
  • Takahata, S., Senju, N., Osaki, Y., Yoshida, T., Ida, T. (2006). Amino Acid Substitutions in Mosaic Penicillin-Binding Protein 2 Associated with Reduced Susceptibility to Cefixime in Clinical Isolates of Neisseria gonorrhoeae. Antimicrob. Agents Chemother. 50: 3638-3645 [Abstract] [Full Text]  
  • Shafer, W. M., Folster, J. P. (2006). Towards an Understanding of Chromosomally Mediated Penicillin Resistance in Neisseria gonorrhoeae: Evidence for a Porin-Efflux Pump Collaboration.. J. Bacteriol. 188: 2297-2299 [Full Text]  
  • Olesky, M., Zhao, S., Rosenberg, R. L., Nicholas, R. A. (2006). Porin-Mediated Antibiotic Resistance in Neisseria gonorrhoeae: Ion, Solute, and Antibiotic Permeation through PIB Proteins with penB Mutations.. J. Bacteriol. 188: 2300-2308 [Abstract] [Full Text]  
  • Hu, M., Nandi, S., Davies, C., Nicholas, R. A. (2005). High-Level Chromosomally Mediated Tetracycline Resistance in Neisseria gonorrhoeae Results from a Point Mutation in the rpsJ Gene Encoding Ribosomal Protein S10 in Combination with the mtrR and penB Resistance Determinants. Antimicrob. Agents Chemother. 49: 4327-4334 [Abstract] [Full Text]  
  • Perez-Losada, M., Viscidi, R. P., Demma, J. C., Zenilman, J., Crandall, K. A. (2005). Population Genetics of Neisseria gonorrhoeae in a High-Prevalence Community Using a Hypervariable Outer Membrane porB and 13 Slowly Evolving Housekeeping Genes. Mol Biol Evol 22: 1887-1902 [Abstract] [Full Text]  
  • Vereshchagin, V. A., Ilina, E. N., Malakhova, M. V., Zubkov, M. M., Sidorenko, S. V., Kubanova, A. A., Govorun, V. M. (2004). Fluoroquinolone-resistant Neisseria gonorrhoeae isolates from Russia: molecular mechanisms implicated. J Antimicrob Chemother 53: 653-656 [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»