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 Payton, M.
Right arrow Articles by Sim, E.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Payton, M.
Right arrow Articles by Sim, E.

 Previous Article  |  Next Article 

Journal of Bacteriology, February 1999, p. 1343-1347, Vol. 181, No. 4
0021-9193/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

Cloning and Characterization of Arylamine N-Acetyltransferase Genes from Mycobacterium smegmatis and Mycobacterium tuberculosis: Increased Expression Results in Isoniazid Resistance

Mark Payton,1 Roy Auty,1 Rupika Delgoda,1 Martin Everett,2 and Edith Sim1,*

Department of Pharmacology, University of Oxford, Oxford OX1 3QT,1 and Research and Development, Glaxo-Wellcome plc, Stevenage, Hertfordshire SG1 2NY,2 United Kingdom

Received 9 September 1998/Accepted 3 December 1998

Arylamine N-acetyltransferases (NATs) are found in many eukaryotic organisms, including humans, and have previously been identified in the prokaryote Salmonella typhimurium. NATs from many sources acetylate the antitubercular drug isoniazid and so inactivate it. nat genes were cloned from Mycobacterium smegmatis and Mycobacterium tuberculosis, and expressed in Escherichia coli and M. smegmatis. The induced M. smegmatis NAT catalyzes the acetylation of isoniazid. A monospecific antiserum raised against pure NAT from S. typhimurium recognizes NAT from M. smegmatis and cross-reacts with recombinant NAT from M. tuberculosis. Overexpression of mycobacterial nat genes in E. coli results in predominantly insoluble recombinant protein; however, with M. smegmatis as the host using the vector pACE-1, NAT proteins from M. tuberculosis and M. smegmatis are soluble. M. smegmatis transformants induced to express the M. tuberculosis nat gene in culture demonstrated a threefold higher resistance to isoniazid. We propose that NAT in mycobacteria could have a role in acetylating, and hence inactivating, isoniazid.

* Corresponding author. Mailing address: Department of Pharmacology, University of Oxford, Mansfield Rd., Oxford OX1 3QT, United Kingdom. Phone: (44 1865) 271596. Fax: (44 1865) 271853. E-mail: esim{at}worf.molbiol.ox.ac.uk.

Journal of Bacteriology, February 1999, p. 1343-1347, Vol. 181, No. 4
0021-9193/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


This article has been cited by other articles:

  • Srivastava, V., Rouanet, C., Srivastava, R., Ramalingam, B., Locht, C., Srivastava, B. S. (2007). Macrophage-specific Mycobacterium tuberculosis genes: identification by green fluorescent protein and kanamycin resistance selection. Microbiology 153: 659-666 [Abstract] [Full Text]  
  • Suzuki, H., Ohnishi, Y., Horinouchi, S. (2007). Arylamine N-Acetyltransferase Responsible for Acetylation of 2-Aminophenols in Streptomyces griseus. J. Bacteriol. 189: 2155-2159 [Abstract] [Full Text]  
  • Adjei, M. D., Heinze, T. M., Deck, J., Freeman, J. P., Williams, A. J., Sutherland, J. B. (2006). Transformation of the Antibacterial Agent Norfloxacin by Environmental Mycobacteria. Appl. Environ. Microbiol. 72: 5790-5793 [Abstract] [Full Text]  
  • Sholto-Douglas-Vernon, C., Sandy, J., Victor, T. C, Sim, E., Helden, P. D. (2005). Mutational and expression analysis of tbnat and its response to isoniazid. J Med Microbiol 54: 1189-1197 [Abstract] [Full Text]  
  • Bhakta, S., Besra, G. S., Upton, A. M., Parish, T., Sholto-Douglas-Vernon, C., Gibson, K. J.C., Knutton, S., Gordon, S., daSilva, R. P., Anderton, M. C., Sim, E. (2004). Arylamine N-Acetyltransferase Is Required for Synthesis of Mycolic Acids and Complex Lipids in Mycobacterium bovis BCG and Represents a Novel Drug Target. JEM 199: 1191-1199 [Abstract] [Full Text]  
  • Hearn, M. J., Cynamon, M. H. (2004). Design and synthesis of antituberculars: preparation and evaluation against Mycobacterium tuberculosis of an isoniazid Schiff base. J Antimicrob Chemother 53: 185-191 [Abstract] [Full Text]  
  • Ramaswamy, S. V., Reich, R., Dou, S.-J., Jasperse, L., Pan, X., Wanger, A., Quitugua, T., Graviss, E. A. (2003). Single Nucleotide Polymorphisms in Genes Associated with Isoniazid Resistance in Mycobacterium tuberculosis. Antimicrob. Agents Chemother. 47: 1241-1250 [Abstract] [Full Text]  
  • Viveiros, M., Portugal, I., Bettencourt, R., Victor, T. C., Jordaan, A. M., Leandro, C., Ordway, D., Amaral, L. (2002). Isoniazid-Induced Transient High-Level Resistance in Mycobacterium tuberculosis. Antimicrob. Agents Chemother. 46: 2804-2810 [Abstract] [Full Text]  
  • Mushtaq, A., Payton, M., Sim, E. (2002). The COOH Terminus of Arylamine N-Acetyltransferase from Salmonella typhimurium Controls Enzymic Activity. J. Biol. Chem. 277: 12175-12181 [Abstract] [Full Text]  
  • Diaz, E., Ferrandez, A., Prieto, M. A., Garcia, J. L. (2001). Biodegradation of Aromatic Compounds by Escherichia coli. Microbiol. Mol. Biol. Rev. 65: 523-569 [Abstract] [Full Text]  
  • Payton, M., Gifford, C., Schartau, P., Hagemeier, C., Mushtaq, A., Lucas, S., Pinter, K., Sim, E. (2001). Evidence towards the role of arylamine N-acetyltransferase in Mycobacterium smegmatis and development of a specific antiserum against the homologous enzyme of Mycobacterium tuberculosis. Microbiology 147: 3295-3302 [Abstract] [Full Text]  
  • Deloménie, C., Fouix, S., Longuemaux, S., Brahimi, N., Bizet, C., Picard, B., Denamur, E., Dupret, J.-M. (2001). Identification and Functional Characterization of Arylamine N-Acetyltransferases in Eubacteria: Evidence for Highly Selective Acetylation of 5-Aminosalicylic Acid. J. Bacteriol. 183: 3417-3427 [Abstract] [Full Text]  
  • Payton, M., Mushtaq, A., Yu, T.-W., Wu, L.-J., Sinclair, J., Sim, E. (2001). Eubacterial arylamine N-acetyltransferases - identification and comparison of 18 members of the protein family with conserved active site cysteine, histidine and aspartate residues. Microbiology 147: 1137-1147 [Abstract] [Full Text]  
  • Sim, E., Payton, M., Noble, M., Minchin, R. (2000). An update on genetic, structural and functional studies of arylamine N-acetyltransferases in eucaryotes and procaryotes. Hum Mol Genet 9: 2435-2441 [Abstract] [Full Text]  
  • Vemulapalli, R., He, Y., Cravero, S., Sriranganathan, N., Boyle, S. M., Schurig, G. G. (2000). Overexpression of Protective Antigen as a Novel Approach To Enhance Vaccine Efficacy of Brucella abortus Strain RB51. Infect. Immun. 68: 3286-3289 [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»