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 Ji, Y.
Right arrow Articles by Woodnutt, G.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Ji, Y.
Right arrow Articles by Woodnutt, G.

 Previous Article  |  Next Article 

Journal of Bacteriology, November 1999, p. 6585-6590, Vol. 181, No. 21
0021-9193/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

Regulated Antisense RNA Eliminates Alpha-Toxin Virulence in Staphylococcus aureus Infection

Yinduo Ji,* Andrea Marra,dagger Martin Rosenberg, and Gary Woodnutt

Department of Microbiology, SmithKline Beecham Pharmaceuticals Research and Development, Collegeville, Pennsylvania 19426

Received 23 June 1999/Accepted 20 August 1999

The ability to selectively disrupt gene function remains a critical element in elucidating information regarding gene essentiality for bacterial growth and/or pathogenesis. In this study, we adapted a tet regulatory expression system for use in Staphylococcus aureus, with the goal of downregulating gene expression via induction of antisense RNA. We demonstrate that this system exhibits a 50- to 100-fold dose-dependent level of induction in bacterial cells grown in culture (i.e., in vitro) and also functions in mice (i.e., in vivo) following oral administration of inducer. To determine whether induced antisense RNA could interfere with chromosomally derived gene expression, we cloned a fragment of the S. aureus alpha-toxin gene (hla) in antisense orientation downstream of the tet promoter system and introduced the construct into S. aureus. Induced antisense hla RNA downregulated chromosomally derived hla gene expression in vitro approximately 14-fold. Similarly, induction of hla antisense RNA in vivo dramatically reduced alpha-toxin expression in two different murine models of S. aureus infection. Most importantly, this reduction completely eliminated the lethality of the infection. These results indicate that the tet regulatory system functions efficiently in S. aureus and induced antisense RNA can effectively downregulate chromosomal gene expression both in vitro and in vivo.

* Corresponding author. Mailing address: 1250 South Collegeville Road, UP1345, Collegeville, PA 19426. Phone: (610) 917-7663. Fax: (610) 917-7901. E-mail: Yinduo_Ji-1{at}sbphrd.com.

dagger Present address: Protein Design Labs, Fremont, CA 94555.

Journal of Bacteriology, November 1999, p. 6585-6590, Vol. 181, No. 21
0021-9193/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


This article has been cited by other articles:

  • Sastalla, I., Chim, K., Cheung, G. Y. C., Pomerantsev, A. P., Leppla, S. H. (2009). Codon-Optimized Fluorescent Proteins Designed for Expression in Low-GC Gram-Positive Bacteria. Appl. Environ. Microbiol. 75: 2099-2110 [Abstract] [Full Text]  
  • Liang, X., Yan, M., Ji, Y. (2009). The H35A Mutated Alpha-Toxin Interferes with Cytotoxicity of Staphylococcal Alpha-Toxin. Infect. Immun. 77: 977-983 [Abstract] [Full Text]  
  • Yarovinsky, T. O., Monick, M. M., Husmann, M., Hunninghake, G. W. (2008). Interferons Increase Cell Resistance to Staphylococcal Alpha-Toxin. Infect. Immun. 76: 571-577 [Abstract] [Full Text]  
  • Kedar, G. C., Brown-Driver, V., Reyes, D. R., Hilgers, M. T., Stidham, M. A., Shaw, K. J., Finn, J., Haselbeck, R. J. (2007). Evaluation of the metS and murB Loci for Antibiotic Discovery Using Targeted Antisense RNA Expression Analysis in Bacillus anthracis. Antimicrob. Agents Chemother. 51: 1708-1718 [Abstract] [Full Text]  
  • Liang, X., Yu, C., Sun, J., Liu, H., Landwehr, C., Holmes, D., Ji, Y. (2006). Inactivation of a Two-Component Signal Transduction System, SaeRS, Eliminates Adherence and Attenuates Virulence of Staphylococcus aureus.. Infect. Immun. 74: 4655-4665 [Abstract] [Full Text]  
  • Dubytska, L., Godfrey, H. P., Cabello, F. C. (2006). Borrelia burgdorferi ftsZ Plays a Role in Cell Division.. J. Bacteriol. 188: 1969-1978 [Abstract] [Full Text]  
  • Pomerantsev, A. P., Sitaraman, R., Galloway, C. R., Kivovich, V., Leppla, S. H. (2006). Genome Engineering in Bacillus anthracis Using Cre Recombinase. Infect. Immun. 74: 682-693 [Abstract] [Full Text]  
  • Sun, J., Zheng, L., Landwehr, C., Yang, J., Ji, Y. (2005). Identification of a Novel Essential Two-Component Signal Transduction System, YhcSR, in Staphylococcus aureus. J. Bacteriol. 187: 7876-7880 [Abstract] [Full Text]  
  • Bertram, R., Kostner, M., Muller, J., Ramos, J. V., Hillen, W. (2005). Integrative elements for Bacillus subtilis yielding tetracycline-dependent growth phenotypes. Nucleic Acids Res 33: e153-e153 [Abstract] [Full Text]  
  • Liang, X., Zheng, L., Landwehr, C., Lunsford, D., Holmes, D., Ji, Y. (2005). Global Regulation of Gene Expression by ArlRS, a Two-Component Signal Transduction Regulatory System of Staphylococcus aureus. J. Bacteriol. 187: 5486-5492 [Abstract] [Full Text]  
  • Wang, B., Kuramitsu, H. K. (2005). Inducible Antisense RNA Expression in the Characterization of Gene Functions in Streptococcus mutans. Infect. Immun. 73: 3568-3576 [Abstract] [Full Text]  
  • Kamionka, A., Bertram, R., Hillen, W. (2005). Tetracycline-Dependent Conditional Gene Knockout in Bacillus subtilis. Appl. Environ. Microbiol. 71: 728-733 [Abstract] [Full Text]  
  • Blokpoel, M. C. J., Murphy, H. N., O'Toole, R., Wiles, S., Runn, E. S. C., Stewart, G. R., Young, D. B., Robertson, B. D. (2005). Tetracycline-inducible gene regulation in mycobacteria. Nucleic Acids Res 33: e22-e22 [Abstract] [Full Text]  
  • Chen, G., Patten, C. L., Schellhorn, H. E. (2003). Controlled Expression of an rpoS Antisense RNA Can Inhibit RpoS Function in Escherichia coli. Antimicrob. Agents Chemother. 47: 3485-3493 [Abstract] [Full Text]  
  • Pomerantsev, A. P., Kalnin, K. V., Osorio, M., Leppla, S. H. (2003). Phosphatidylcholine-Specific Phospholipase C and Sphingomyelinase Activities in Bacteria of the Bacillus cereus Group. Infect. Immun. 71: 6591-6606 [Abstract] [Full Text]  
  • Weichhart, T., Horky, M., Sollner, J., Gangl, S., Henics, T., Nagy, E., Meinke, A., von Gabain, A., Fraser, C. M., Gill, S. R., Hafner, M., von Ahsen, U. (2003). Functional Selection of Vaccine Candidate Peptides from Staphylococcus aureus Whole-Genome Expression Libraries In Vitro. Infect. Immun. 71: 4633-4641 [Abstract] [Full Text]  
  • Schofield, D. A., Westwater, C., Hoel, B. D., Werner, P. A., Norris, J. S., Schmidt, M. G. (2003). Development of a Thermally Regulated Broad-Spectrum Promoter System for Use in Pathogenic Gram-Positive Species. Appl. Environ. Microbiol. 69: 3385-3392 [Abstract] [Full Text]  
  • Bateman, B. T., Donegan, N. P., Jarry, T. M., Palma, M., Cheung, A. L. (2001). Evaluation of a Tetracycline-Inducible Promoter in Staphylococcus aureus In Vitro and In Vivo and Its Application in Demonstrating the Role of sigB in Microcolony Formation. Infect. Immun. 69: 7851-7857 [Abstract] [Full Text]  
  • Kielian, T., Cheung, A., Hickey, W. F. (2001). Diminished Virulence of an Alpha-Toxin Mutant of Staphylococcus aureus in Experimental Brain Abscesses. Infect. Immun. 69: 6902-6911 [Abstract] [Full Text]  
  • Ji, Y., Zhang, B., Van, S. F., Horn, , Warren, P., Woodnutt, G., Burnham, M. K. R., Rosenberg, M. (2001). Identification of Critical Staphylococcal Genes Using Conditional Phenotypes Generated by Antisense RNA. Science 293: 2266-2269 [Abstract] [Full Text]  
  • Morrissey, J. A., Cockayne, A., Hill, P. J., Williams, P. (2000). Molecular Cloning and Analysis of a Putative Siderophore ABC Transporter from Staphylococcus aureus. Infect. Immun. 68: 6281-6288 [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»