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 Rice, L. B.
Right arrow Articles by Bonomo, R. A.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Rice, L. B.
Right arrow Articles by Bonomo, R. A.

 Previous Article  |  Next Article 

Antimicrobial Agents and Chemotherapy, February 2000, p. 362-367, Vol. 44, No. 2
0066-4804/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

High-Level Expression of Chromosomally Encoded SHV-1 beta -Lactamase and an Outer Membrane Protein Change Confer Resistance to Ceftazidime and Piperacillin- Tazobactam in a Clinical Isolate of Klebsiella pneumoniae

Louis B. Rice,1,2,* Lenore L. Carias,2 Andrea M. Hujer,3 Mary Bonafede,4 Rebecca Hutton,3 Claudia Hoyen,4 and Robert A. Bonomo1,2

Medical1 and Research3 Services, Department of Veterans Affairs Medical Center, and Departments of Medicine2 and Pediatrics,4 Case Western Reserve University School of Medicine, Cleveland, Ohio

Received 15 March 1999/Returned for modification 23 June 1999/Accepted 9 November 1999

We describe Klebsiella pneumoniae 15571, a clinical isolate resistant to ceftazidime MIC = 32 µg/ml) and piperacillin-tazobactam (MICs = 1,024 and 128 µg/ml). K. pneumoniae 15571 expresses a single beta -lactamase with a pI of 7.6. However, when cloned in a high-copy-number vector in Escherichia coli, this blaSHV-1 gene did not confer resistance to ceftazidime, a spectrum consistent with the nucleotide sequence, which was nearly identical to those of previously described blaSHV-1 genes. Outer membrane protein (OMP) analysis of K. pneumoniae 15571 revealed a decrease in the quantity of a minor 45-kDa OMP in comparison to that in K. pneumoniae 44NR, a low-level ampicillin-resistant strain that also expresses a chromosomally determined blaSHV-1. Crude beta -lactamase enzyme extracts from K. pneumoniae 15571 produced roughly 200-fold more beta -lactamase activity than K. pneumoniae 44NR. Northern hybridization analysis revealed that this difference was explainable by quantifiable differences in transcription of the blaSHV-1 gene in the two strains. Primer extension analysis of blaSHV-1 mRNA from K. pneumoniae 15571 and 44NR indicated that the transcriptional start sites were identical in the two strains. DNA sequencing of the promoter regions upstream of the of blaSHV-1 open reading frames in the two K. pneumoniae strains revealed an Aright-arrowC change in the second position of the -10 region in K. pneumoniae 44NR compared to that in 15571. Site-directed mutagenesis of the cloned K. pneumoniae 15571 blaSHV-1, in which the A in the second position of the 15571 -10 region was changed to a C, resulted in a substantial lowering of the MIC of ampicillin. When the levels of beta -lactamase enzyme expression in E. coli were compared, the blaSHV-1 downstream of the altered -10 region produced 17-fold less beta -lactamase enzyme. These results indicate that elevated levels of ceftazidime resistance can result from a combination of increased enzyme production and minor OMP changes and that levels of chromosomally encoded SHV-1 beta -lactamase production can vary substantially with a single-base-pair change in promoter sequence.

* Corresponding author. Mailing address: Medical Service 111(W), VA Medical Center, 10701 East Blvd., Cleveland, OH 44106. Phone: (216) 791-3800, ext. 4800. Fax: (216) 231-3289. E-mail: louis.rice{at}med.va.gov.

Antimicrobial Agents and Chemotherapy, February 2000, p. 362-367, Vol. 44, No. 2
0066-4804/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


This article has been cited by other articles:

  • Turner, M. S., Andersson, P., Bell, J. M., Turnidge, J. D., Harris, T., Giffard, P. M. (2009). Plasmid-borne blaSHV genes in Klebsiella pneumoniae are associated with strong promoters. J Antimicrob Chemother 64: 960-964 [Abstract] [Full Text]  
  • Pattanaik, P., Bethel, C. R., Hujer, A. M., Hujer, K. M., Distler, A. M., Taracila, M., Anderson, V. E., Fritsche, T. R., Jones, R. N., Pagadala, S. R. R., van den Akker, F., Buynak, J. D., Bonomo, R. A. (2009). Strategic Design of an Effective {beta}-Lactamase Inhibitor: LN-1-255, A 6-ALKYLIDENE-2'-SUBSTITUTED PENICILLIN SULFONE. J. Biol. Chem. 284: 945-953 [Abstract] [Full Text]  
  • Dubois, V., Poirel, L., Demarthe, F., Arpin, C., Coulange, L., Minarini, L. A. R., Bezian, M.-C., Nordmann, P., Quentin, C. (2008). Molecular and Biochemical Characterization of SHV-56, a Novel Inhibitor-Resistant {beta}-Lactamase from Klebsiella pneumoniae. Antimicrob. Agents Chemother. 52: 3792-3794 [Abstract] [Full Text]  
  • Yang, K., Guglielmo, B J. (2007). Diagnosis and Treatment of Extended-Spectrum and AmpC {beta}-Lactamase-Producing Organisms. The Annals of Pharmacotherapy 41: 1427-1435 [Abstract] [Full Text]  
  • Thomson, J. M., Prati, F., Bethel, C. R., Bonomo, R. A. (2007). Use of Novel Boronic Acid Transition State Inhibitors To Probe Substrate Affinity in SHV-Type Extended-Spectrum {beta}-Lactamases. Antimicrob. Agents Chemother. 51: 1577-1579 [Abstract] [Full Text]  
  • Tofteland, S., Haldorsen, B., Dahl, K. H., Simonsen, G. S., Steinbakk, M., Walsh, T. R., Sundsfjord, A., the Norwegian ESBL Study Group, (2007). Effects of Phenotype and Genotype on Methods for Detection of Extended-Spectrum-{beta}-Lactamase-Producing Clinical Isolates of Escherichia coli and Klebsiella pneumoniae in Norway. J. Clin. Microbiol. 45: 199-205 [Abstract] [Full Text]  
  • Bethel, C. R., Hujer, A. M., Hujer, K. M., Thomson, J. M., Ruszczycky, M. W., Anderson, V. E., Pusztai-Carey, M., Taracila, M., Helfand, M. S., Bonomo, R. A. (2006). Role of Asp104 in the SHV {beta}-Lactamase. Antimicrob. Agents Chemother. 50: 4124-4131 [Abstract] [Full Text]  
  • Hujer, K. M., Hujer, A. M., Hulten, E. A., Bajaksouzian, S., Adams, J. M., Donskey, C. J., Ecker, D. J., Massire, C., Eshoo, M. W., Sampath, R., Thomson, J. M., Rather, P. N., Craft, D. W., Fishbain, J. T., Ewell, A. J., Jacobs, M. R., Paterson, D. L., Bonomo, R. A. (2006). Analysis of Antibiotic Resistance Genes in Multidrug-Resistant Acinetobacter sp. Isolates from Military and Civilian Patients Treated at the Walter Reed Army Medical Center. Antimicrob. Agents Chemother. 50: 4114-4123 [Abstract] [Full Text]  
  • Thomson, J. M., Distler, A. M., Prati, F., Bonomo, R. A. (2006). Probing Active Site Chemistry in SHV beta-Lactamase Variants at Ambler Position 244: UNDERSTANDING UNIQUE PROPERTIES OF INHIBITOR RESISTANCE. J. Biol. Chem. 281: 26734-26744 [Abstract] [Full Text]  
  • Reynolds, K. A., Thomson, J. M., Corbett, K. D., Bethel, C. R., Berger, J. M., Kirsch, J. F., Bonomo, R. A., Handel, T. M. (2006). Structural and Computational Characterization of the SHV-1 beta-Lactamase-beta-Lactamase Inhibitor Protein Interface. J. Biol. Chem. 281: 26745-26753 [Abstract] [Full Text]  
  • Spanu, T., Sanguinetti, M., Tumbarello, M., D'Inzeo, T., Fiori, B., Posteraro, B., Santangelo, R., Cauda, R., Fadda, G. (2006). Evaluation of the New VITEK 2 Extended-Spectrum Beta-Lactamase (ESBL) Test for Rapid Detection of ESBL Production in Enterobacteriaceae Isolates.. J. Clin. Microbiol. 44: 3257-3262 [Abstract] [Full Text]  
  • Paterson, D. L., Bonomo, R. A. (2005). Extended-Spectrum {beta}-Lactamases: a Clinical Update. Clin. Microbiol. Rev. 18: 657-686 [Abstract] [Full Text]  
  • Furushita, M., Okamoto, A., Maeda, T., Ohta, M., Shiba, T. (2005). Isolation of Multidrug-Resistant Stenotrophomonas maltophilia from Cultured Yellowtail (Seriola quinqueradiata) from a Marine Fish Farm. Appl. Environ. Microbiol. 71: 5598-5600 [Abstract] [Full Text]  
  • Siebor, E., Pechinot, A., Duez, J.-M., Neuwirth, C. (2005). One New LEN Enzyme and Two New OKP Enzymes in Klebsiella pneumoniae Clinical Isolates and Proposed Nomenclature for Chromosomal {beta}-Lactamases of This Species. Antimicrob. Agents Chemother. 49: 3097-3098 [Full Text]  
  • Fang, H., Lundberg, C., Olsson-Liljequist, B., Hedin, G., Lindback, E., Rosenberg, A., Struwe, J. (2004). Molecular Epidemiological Analysis of Escherichia coli Isolates Producing Extended-Spectrum {beta}-Lactamases for Identification of Nosocomial Outbreaks in Stockholm, Sweden. J. Clin. Microbiol. 42: 5917-5920 [Abstract] [Full Text]  
  • Dubois, V., Poirel, L., Arpin, C., Coulange, L., Bebear, C., Nordmann, P., Quentin, C. (2004). SHV-49, a Novel Inhibitor-Resistant {beta}-Lactamase in a Clinical Isolate of Klebsiella pneumoniae. Antimicrob. Agents Chemother. 48: 4466-4469 [Abstract] [Full Text]  
  • Walckenaer, E., Poirel, L., Leflon-Guibout, V., Nordmann, P., Nicolas-Chanoine, M.-H. (2004). Genetic and Biochemical Characterization of the Chromosomal Class A {beta}-Lactamases of Raoultella (formerly Klebsiella) planticola and Raoultella ornithinolytica. Antimicrob. Agents Chemother. 48: 305-312 [Abstract] [Full Text]  
  • Helfand, M. S., Bethel, C. R., Hujer, A. M., Hujer, K. M., Anderson, V. E., Bonomo, R. A. (2003). Understanding Resistance to {beta}-Lactams and {beta}-Lactamase Inhibitors in the SHV {beta}-Lactamase: LESSONS FROM THE MUTAGENESIS OF SER-130. J. Biol. Chem. 278: 52724-52729 [Abstract] [Full Text]  
  • Babini, G. S., Yuan, M., Hall, L. M. C., Livermore, D. M. (2003). Variable susceptibility to piperacillin/tazobactam amongst Klebsiella spp. with extended-spectrum {beta}-lactamases. J Antimicrob Chemother 51: 605-612 [Abstract] [Full Text]  
  • Hujer, A. M., Hujer, K. M., Helfand, M. S., Anderson, V. E., Bonomo, R. A. (2002). Amino Acid Substitutions at Ambler Position Gly238 in the SHV-1 {beta}-Lactamase: Exploring Sequence Requirements for Resistance to Penicillins and Cephalosporins. Antimicrob. Agents Chemother. 46: 3971-3977 [Abstract] [Full Text]  
  • Corvec, S., Caroff, N., Espaze, E., Marraillac, J., Reynaud, A. (2002). -11 Mutation in the ampC Promoter Increasing Resistance to {beta}-Lactams in a Clinical Escherichia coli Strain. Antimicrob. Agents Chemother. 46: 3265-3267 [Abstract] [Full Text]  
  • Hujer, A. M., Page, M. G. P., Helfand, M. S., Yeiser, B., Bonomo, R. A. (2002). Development of a Sensitive and Specific Enzyme-Linked Immunosorbent Assay for Detecting and Quantifying CMY-2 and SHV {beta}-Lactamases. J. Clin. Microbiol. 40: 1947-1957 [Abstract] [Full Text]  
  • Howard, C., van Daal, A., Kelly, G., Schooneveldt, J., Nimmo, G., Giffard, P. M. (2002). Identification and Minisequencing-Based Discrimination of SHV {beta}-Lactamases in Nosocomial Infection-Associated Klebsiella pneumoniae in Brisbane, Australia. Antimicrob. Agents Chemother. 46: 659-664 [Abstract] [Full Text]  
  • Bradford, P. A. (2001). Extended-Spectrum {beta}-Lactamases in the 21st Century: Characterization, Epidemiology, and Detection of This Important Resistance Threat. Clin. Microbiol. Rev. 14: 933-951 [Abstract] [Full Text]  
  • Chaves, J., Ladona, M. G., Segura, C., Coira, A., Reig, R., Ampurdanes, C. (2001). SHV-1 {beta}-Lactamase Is Mainly a Chromosomally Encoded Species-Specific Enzyme in Klebsiella pneumoniae. Antimicrob. Agents Chemother. 45: 2856-2861 [Abstract] [Full Text]  
  • Arpin, C., Labia, R., Andre, C., Frigo, C., El Harrif, Z., Quentin, C. (2001). SHV-16, a {beta}-Lactamase with a Pentapeptide Duplication in the Omega Loop. Antimicrob. Agents Chemother. 45: 2480-2485 [Abstract] [Full Text]  
  • Steward, C. D., Rasheed, J. K., Hubert, S. K., Biddle, J. W., Raney, P. M., Anderson, G. J., Williams, P. P., Brittain, K. L., Oliver, A., McGowan, J. E. Jr., Tenover, F. C. (2001). Characterization of Clinical Isolates of Klebsiella pneumoniae from 19 Laboratories Using the National Committee for Clinical Laboratory Standards Extended-Spectrum {beta}-Lactamase Detection Methods. J. Clin. Microbiol. 39: 2864-2872 [Abstract] [Full Text]  
  • Wu, T.-L., Siu, L. K., Su, L.-H., Lauderdale, T. L., Lin, F. M., Leu, H.-S., Lin, T.-Y., Ho, M. (2001). Outer membrane protein change combined with co-existing TEM-1 and SHV-1 {beta}-lactamases lead to false identification of ESBL-producing Klebsiella pneumoniae. J Antimicrob Chemother 47: 755-761 [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»