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 Nizet, V.
Right arrow Articles by De Azavedo, J. C. S.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Nizet, V.
Right arrow Articles by De Azavedo, J. C. S.

 Previous Article  |  Next Article 

Infection and Immunity, July 2000, p. 4245-4254, Vol. 68, No. 7
0019-9567/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Genetic Locus for Streptolysin S Production by Group A Streptococcus

Victor Nizet,1 Bernard Beall,2 Darrin J. Bast,3 Vivekananda Datta,1 Laurie Kilburn,3 Donald E. Low,3 and Joyce C. S. De Azavedo3,*

Department of Pediatrics, Division of Infectious Diseases, University of California, San Diego, La Jolla, California 920931; Centers for Disease Control and Prevention, National Center for Infectious Diseases, Respiratory Diseases Branch, Atlanta, Georgia 303332; and Department of Microbiology, Mount Sinai Hospital and University of Toronto, Toronto, Ontario, Canada M5G 1X53

Received 17 February 2000/Returned for modification 9 March 2000/Accepted 23 March 2000

Group A streptococcus (GAS) is an important human pathogen that causes pharyngitis and invasive infections, including necrotizing fasciitis. Streptolysin S (SLS) is the cytolytic factor that creates the zone of beta-hemolysis surrounding GAS colonies grown on blood agar. We recently reported the discovery of a potential genetic determinant involved in SLS production, sagA, encoding a small peptide of 53 amino acids (S. D. Betschel, S. M. Borgia, N. L. Barg, D. E. Low, and J. C. De Azavedo, Infect. Immun. 66:1671-1679, 1998). Using transposon mutagenesis, chromosomal walking steps, and data from the GAS genome sequencing project (www.genome.ou.edu/strep.html), we have now identified a contiguous nine-gene locus (sagA to sagI) involved in SLS production. The sag locus is conserved among GAS strains regardless of M protein type. Targeted plasmid integrational mutagenesis of each gene in the sag operon resulted in an SLS-negative phenotype. Targeted integrations (i) upstream of the sagA promoter and (ii) downstream of a terminator sequence after sagI did not affect SLS production, establishing the functional boundaries of the operon. A rho-independent terminator sequence between sagA and sagB appears to regulate the amount of sagA transcript produced versus transcript for the entire operon. Reintroduction of the nine-gene sag locus on a plasmid vector restored SLS activity to the nonhemolytic sagA knockout mutant. Finally, heterologous expression of the intact sag operon conferred the SLS beta-hemolytic phenotype to the nonhemolytic Lactococcus lactis. We conclude that gene products of the GAS sag operon are both necessary and sufficient for SLS production. Sequence homologies of sag operon gene products suggest that SLS is related to the bacteriocin family of microbial toxins.


* Corresponding author. Mailing address: Department of Microbiology, Room 1483, Mount Sinai Hospital, 600 University Ave., Toronto, Canada M5G 1X5. Phone: (416) 586-8459. Fax: (416) 586-8746. E-mail: jdeazavedo{at}mtsinai.on.ca.


Infection and Immunity, July 2000, p. 4245-4254, Vol. 68, No. 7
0019-9567/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.



This article has been cited by other articles:

  • Mitchell, D. A., Lee, S. W., Pence, M. A., Markley, A. L., Limm, J. D., Nizet, V., Dixon, J. E. (2009). Structural and Functional Dissection of the Heterocyclic Peptide Cytotoxin Streptolysin S. J. Biol. Chem. 284: 13004-13012 [Abstract] [Full Text]  
  • McShan, W. M., Ferretti, J. J., Karasawa, T., Suvorov, A. N., Lin, S., Qin, B., Jia, H., Kenton, S., Najar, F., Wu, H., Scott, J., Roe, B. A., Savic, D. J. (2008). Genome Sequence of a Nephritogenic and Highly Transformable M49 Strain of Streptococcus pyogenes. J. Bacteriol. 190: 7773-7785 [Abstract] [Full Text]  
  • Lee, S. W., Mitchell, D. A., Markley, A. L., Hensler, M. E., Gonzalez, D., Wohlrab, A., Dorrestein, P. C., Nizet, V., Dixon, J. E. (2008). From the Cover: Discovery of a widely distributed toxin biosynthetic gene cluster. Proc. Natl. Acad. Sci. USA 105: 5879-5884 [Abstract] [Full Text]  
  • Walsh, C. T., Nolan, E. M. (2008). Morphing peptide backbones into heterocycles. Proc. Natl. Acad. Sci. USA 105: 5655-5656 [Full Text]  
  • Shelburne, S. A. III, Keith, D., Horstmann, N., Sumby, P., Davenport, M. T., Graviss, E. A., Brennan, R. G., Musser, J. M. (2008). A direct link between carbohydrate utilization and virulence in the major human pathogen group A Streptococcus. Proc. Natl. Acad. Sci. USA 105: 1698-1703 [Abstract] [Full Text]  
  • Salim, K. Y., de Azavedo, J. C., Bast, D. J., Cvitkovitch, D. G. (2007). Role for sagA and siaA in Quorum Sensing and Iron Regulation in Streptococcus pyogenes. Infect. Immun. 75: 5011-5017 [Abstract] [Full Text]  
  • Nes, I. F., Diep, D. B., Holo, H. (2007). Bacteriocin Diversity in Streptococcus and Enterococcus. J. Bacteriol. 189: 1189-1198 [Full Text]  
  • Kwinn, L. A., Khosravi, A., Aziz, R. K., Timmer, A. M., Doran, K. S., Kotb, M., Nizet, V. (2007). Genetic Characterization and Virulence Role of the RALP3/LSA Locus Upstream of the Streptolysin S Operon in Invasive M1T1 Group A Streptococcus. J. Bacteriol. 189: 1322-1329 [Abstract] [Full Text]  
  • Marri, P. R., Hao, W., Golding, G. B. (2006). Gene Gain and Gene Loss in Streptococcus: Is It Driven by Habitat?. Mol Biol Evol 23: 2379-2391 [Abstract] [Full Text]  
  • Lyon, W. R., Caparon, M. G. (2004). Role for Serine Protease HtrA (DegP) of Streptococcus pyogenes in the Biogenesis of Virulence Factors SpeB and the Hemolysin Streptolysin S. Infect. Immun. 72: 1618-1625 [Abstract] [Full Text]  
  • Engleberg, N. C., Heath, A., Vardaman, K., DiRita, V. J. (2004). Contribution of CsrR-Regulated Virulence Factors to the Progress and Outcome of Murine Skin Infections by Streptococcus pyogenes. Infect. Immun. 72: 623-628 [Abstract] [Full Text]  
  • Fontaine, M. C., Lee, J. J., Kehoe, M. A. (2003). Combined Contributions of Streptolysin O and Streptolysin S to Virulence of Serotype M5 Streptococcus pyogenes Strain Manfredo. Infect. Immun. 71: 3857-3865 [Abstract] [Full Text]  
  • Wescombe, P. A., Tagg, J. R. (2003). Purification and Characterization of Streptin, a Type A1 Lantibiotic Produced by Streptococcus pyogenes. Appl. Environ. Microbiol. 69: 2737-2747 [Abstract] [Full Text]  
  • Gryllos, I., Levin, J. C., Wessels, M. R. (2003). The CsrR/CsrS two-component system of group A Streptococcus responds to environmental Mg2+. Proc. Natl. Acad. Sci. USA 100: 4227-4232 [Abstract] [Full Text]  
  • Sierig, G., Cywes, C., Wessels, M. R., Ashbaugh, C. D. (2003). Cytotoxic Effects of Streptolysin O and Streptolysin S Enhance the Virulence of Poorly Encapsulated Group A Streptococci. Infect. Immun. 71: 446-455 [Abstract] [Full Text]  
  • Fuller, J. D., Camus, A. C., Duncan, C. L., Nizet, V., Bast, D. J., Thune, R. L., Low, D. E., de Azavedo, J. C. S. (2002). Identification of a Streptolysin S-Associated Gene Cluster and Its Role in the Pathogenesis of Streptococcus iniae Disease. Infect. Immun. 70: 5730-5739 [Abstract] [Full Text]  
  • Steiner, K., Malke, H. (2002). Dual Control of Streptokinase and Streptolysin S Production by the covRS and fasCAX Two-Component Regulators in Streptococcus dysgalactiae subsp. equisimilis. Infect. Immun. 70: 3627-3636 [Abstract] [Full Text]  
  • Dale, J. B., Chiang, E. Y., Hasty, D. L., Courtney, H. S. (2002). Antibodies against a Synthetic Peptide of SagA Neutralize the Cytolytic Activity of Streptolysin S from Group A Streptococci. Infect. Immun. 70: 2166-2170 [Abstract] [Full Text]  
  • Gibson, C. M., Caparon, M. G. (2002). Alkaline Phosphatase Reporter Transposon for Identification of Genes Encoding Secreted Proteins in Gram-Positive Microorganisms. Appl. Environ. Microbiol. 68: 928-932 [Abstract] [Full Text]  
  • Steiner, K., Malke, H. (2001). relA-Independent Amino Acid Starvation Response Network of Streptococcus pyogenes. J. Bacteriol. 183: 7354-7364 [Abstract] [Full Text]  
  • Carr, A., Sledjeski, D. D., Podbielski, A., Boyle, M. D. P., Kreikemeyer, B. (2001). Similarities between Complement-mediated and Streptolysin S-mediated Hemolysis. J. Biol. Chem. 276: 41790-41796 [Abstract] [Full Text]  
  • Biswas, I., Germon, P., McDade, K., Scott, J. R. (2001). Generation and Surface Localization of Intact M Protein in Streptococcus pyogenes Are Dependent on sagA. Infect. Immun. 69: 7029-7038 [Abstract] [Full Text]  
  • Upton, M., Tagg, J. R., Wescombe, P., Jenkinson, H. F. (2001). Intra- and Interspecies Signaling between Streptococcus salivarius and Streptococcus pyogenes Mediated by SalA and SalA1 Lantibiotic Peptides. J. Bacteriol. 183: 3931-3938 [Abstract] [Full Text]  
  • Ferretti, J. J., McShan, W. M., Ajdic, D., Savic, D. J., Savic, G., Lyon, K., Primeaux, C., Sezate, S., Suvorov, A. N., Kenton, S., Lai, H. S., Lin, S. P., Qian, Y., Jia, H. G., Najar, F. Z., Ren, Q., Zhu, H., Song, L., White, J., Yuan, X., Clifton, S. W., Roe, B. A., McLaughlin, R. (2001). Complete genome sequence of an M1 strain of Streptococcus pyogenes. Proc. Natl. Acad. Sci. USA 98: 4658-4663 [Abstract] [Full Text]