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. 1995 Jan;63(1):9–20. doi: 10.1128/iai.63.1.9-20.1995

The group A streptococcal virR49 gene controls expression of four structural vir regulon genes.

A Podbielski 1, A Flosdorff 1, J Weber-Heynemann 1
PMCID: PMC172951  PMID: 7806389

Abstract

Within a genomic locus termed the vir regulon, virR genes of opacity factor-nonproducing (OF-) group A streptococci (GAS) are known to control the expression of the genes encoding M protein (emm) and C5a peptidase (scpA) and of virR itself. Within the corresponding genomic locus, opacity factor-producing (OF+) GAS harbor additional emm-related genes encoding immunoglobulin G- and immunoglobulin A-binding proteins (fcrA and enn, respectively). The virR gene region of the OF+ GAS M-type 49 strain CS101 was amplified by PCR, and 2,650 bp were directly sequenced. An open reading frame of 1,599 bp exhibited 76% overall homology to published virR sequences. By utilizing mRNA analysis, the 5' ends of two specific transcripts were mapped 370 and 174 bp upstream of the start codon of this open reading frame. The deduced sequences of the corresponding promoters and their locations differed from those of previously reported virR promoters. Transcripts from wild-type fcrA49, emm49, enn49, and scpA49 genes located downstream of virR49 were characterized as being monocistronic. The transcripts were quantified and mapped for their 5' ends. Subsequently, the virR49 gene was inactivated by specific insertion of a nonreplicative pSF152 vector containing recombinant virR49 sequences. The RNA from the resulting vir-mut strain did not contain transcripts of virR49, fcrA49, emm49, or enn49 and contained reduced amounts of the scpA49 transcript when compared with wild-type RNA. The mRNA control from the streptokinase gene was demonstrated not to be affected. When strain vir-mut was rotated in human blood, it was found to be fully sensitive to phagocytosis by human leukocytes. Thus, the present study provides evidence that virR genes in OF+ GAS could be involved in the control of up to five vir regulon genes, and their unaffected regulatory activity is associated with features postulated as crucial for GAS virulence.

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Selected References

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  1. Bessen D. E., Fischetti V. A. Differentiation between two biologically distinct classes of group A streptococci by limited substitutions of amino acids within the shared region of M protein-like molecules. J Exp Med. 1990 Dec 1;172(6):1757–1764. doi: 10.1084/jem.172.6.1757. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bessen D. E., Fischetti V. A. Nucleotide sequences of two adjacent M or M-like protein genes of group A streptococci: different RNA transcript levels and identification of a unique immunoglobulin A-binding protein. Infect Immun. 1992 Jan;60(1):124–135. doi: 10.1128/iai.60.1.124-135.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bessen D., Fischetti V. A. A human IgG receptor of group A streptococci is associated with tissue site of infection and streptococcal class. J Infect Dis. 1990 Apr;161(4):747–754. doi: 10.1093/infdis/161.4.747. [DOI] [PubMed] [Google Scholar]
  4. Caparon M. G., Geist R. T., Perez-Casal J., Scott J. R. Environmental regulation of virulence in group A streptococci: transcription of the gene encoding M protein is stimulated by carbon dioxide. J Bacteriol. 1992 Sep;174(17):5693–5701. doi: 10.1128/jb.174.17.5693-5701.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Caparon M. G., Scott J. R. Genetic manipulation of pathogenic streptococci. Methods Enzymol. 1991;204:556–586. doi: 10.1016/0076-6879(91)04028-m. [DOI] [PubMed] [Google Scholar]
  6. Caparon M. G., Scott J. R. Identification of a gene that regulates expression of M protein, the major virulence determinant of group A streptococci. Proc Natl Acad Sci U S A. 1987 Dec;84(23):8677–8681. doi: 10.1073/pnas.84.23.8677. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Chen C. C., Cleary P. P. Complete nucleotide sequence of the streptococcal C5a peptidase gene of Streptococcus pyogenes. J Biol Chem. 1990 Feb 25;265(6):3161–3167. [PubMed] [Google Scholar]
  8. Chen C., Bormann N., Cleary P. P. VirR and Mry are homologous trans-acting regulators of M protein and C5a peptidase expression in group A streptococci. Mol Gen Genet. 1993 Dec;241(5-6):685–693. doi: 10.1007/BF00279912. [DOI] [PubMed] [Google Scholar]
  9. Chomczynski P. Solubilization in formamide protects RNA from degradation. Nucleic Acids Res. 1992 Jul 25;20(14):3791–3792. doi: 10.1093/nar/20.14.3791. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Cleuziat P., Awadé A., Robert-Baudouy J. Molecular characterization of pcp, the structural gene encoding the pyrrolidone carboxylyl peptidase from Streptococcus pyogenes. Mol Microbiol. 1992 Aug;6(15):2051–2063. doi: 10.1111/j.1365-2958.1992.tb01378.x. [DOI] [PubMed] [Google Scholar]
  11. Colman G., Tanna A., Efstratiou A., Gaworzewska E. T. The serotypes of Streptococcus pyogenes present in Britain during 1980-1990 and their association with disease. J Med Microbiol. 1993 Sep;39(3):165–178. doi: 10.1099/00222615-39-3-165. [DOI] [PubMed] [Google Scholar]
  12. Coote J. G. Antigenic switching and pathogenicity: environmental effects on virulence gene expression in Bordetella pertussis. J Gen Microbiol. 1991 Nov;137(11):2493–2503. doi: 10.1099/00221287-137-11-2493. [DOI] [PubMed] [Google Scholar]
  13. Erlich H. A., Gelfand D., Sninsky J. J. Recent advances in the polymerase chain reaction. Science. 1991 Jun 21;252(5013):1643–1651. doi: 10.1126/science.2047872. [DOI] [PubMed] [Google Scholar]
  14. Geist R. T., Okada N., Caparon M. G. Analysis of Streptococcus pyogenes promoters by using novel Tn916-based shuttle vectors for the construction of transcriptional fusions to chloramphenicol acetyltransferase. J Bacteriol. 1993 Dec;175(23):7561–7570. doi: 10.1128/jb.175.23.7561-7570.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Haanes-Fritz E., Kraus W., Burdett V., Dale J. B., Beachey E. H., Cleary P. Comparison of the leader sequences of four group A streptococcal M protein genes. Nucleic Acids Res. 1988 May 25;16(10):4667–4677. doi: 10.1093/nar/16.10.4667. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Haanes E. J., Cleary P. P. Identification of a divergent M protein gene and an M protein-related gene family in Streptococcus pyogenes serotype 49. J Bacteriol. 1989 Dec;171(12):6397–6408. doi: 10.1128/jb.171.12.6397-6408.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Haanes E. J., Heath D. G., Cleary P. P. Architecture of the vir regulons of group A streptococci parallels opacity factor phenotype and M protein class. J Bacteriol. 1992 Aug;174(15):4967–4976. doi: 10.1128/jb.174.15.4967-4976.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Heath D. G., Boyle M. D., Cleary P. P. Isolated DNA repeat region from fcrA76, the Fc-binding protein gene from an M-type 76 strain of group A streptococci, encodes a protein with Fc-binding activity. Mol Microbiol. 1990 Dec;4(12):2071–2079. doi: 10.1111/j.1365-2958.1990.tb00567.x. [DOI] [PubMed] [Google Scholar]
  19. Heath D. G., Cleary P. P. Fc-receptor and M-protein genes of group A streptococci are products of gene duplication. Proc Natl Acad Sci U S A. 1989 Jun;86(12):4741–4745. doi: 10.1073/pnas.86.12.4741. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Hollingshead S. K., Readdy T. L., Yung D. L., Bessen D. E. Structural heterogeneity of the emm gene cluster in group A streptococci. Mol Microbiol. 1993 May;8(4):707–717. doi: 10.1111/j.1365-2958.1993.tb01614.x. [DOI] [PubMed] [Google Scholar]
  21. Huang T. T., Malke H., Ferretti J. J. Heterogeneity of the streptokinase gene in group A streptococci. Infect Immun. 1989 Feb;57(2):502–506. doi: 10.1128/iai.57.2.502-506.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Johnson D. R., Stevens D. L., Kaplan E. L. Epidemiologic analysis of group A streptococcal serotypes associated with severe systemic infections, rheumatic fever, or uncomplicated pharyngitis. J Infect Dis. 1992 Aug;166(2):374–382. doi: 10.1093/infdis/166.2.374. [DOI] [PubMed] [Google Scholar]
  23. Kaufhold A., Podbielski A., Johnson D. R., Kaplan E. L., Lütticken R. M protein gene typing of Streptococcus pyogenes by nonradioactively labeled oligonucleotide probes. J Clin Microbiol. 1992 Sep;30(9):2391–2397. doi: 10.1128/jcm.30.9.2391-2397.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Kehoe M. A., Miller L., Walker J. A., Boulnois G. J. Nucleotide sequence of the streptolysin O (SLO) gene: structural homologies between SLO and other membrane-damaging, thiol-activated toxins. Infect Immun. 1987 Dec;55(12):3228–3232. doi: 10.1128/iai.55.12.3228-3232.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. LANCEFIELD R. C. Differentiation of group A streptococci with a common R antigen into three serological types, with special reference to the bactericidal test. J Exp Med. 1957 Oct 1;106(4):525–544. doi: 10.1084/jem.106.4.525. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. LeBlanc D. J., Lee L. N., Inamine J. M. Cloning and nucleotide base sequence analysis of a spectinomycin adenyltransferase AAD(9) determinant from Enterococcus faecalis. Antimicrob Agents Chemother. 1991 Sep;35(9):1804–1810. doi: 10.1128/aac.35.9.1804. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Lottenberg R., Broder C. C., Boyle M. D., Kain S. J., Schroeder B. L., Curtiss R., 3rd Cloning, sequence analysis, and expression in Escherichia coli of a streptococcal plasmin receptor. J Bacteriol. 1992 Aug;174(16):5204–5210. doi: 10.1128/jb.174.16.5204-5210.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Martin N. J., Kaplan E. L., Gerber M. A., Menegus M. A., Randolph M., Bell K., Cleary P. P. Comparison of epidemic and endemic group G streptococci by restriction enzyme analysis. J Clin Microbiol. 1990 Sep;28(9):1881–1886. doi: 10.1128/jcm.28.9.1881-1886.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Mekalanos J. J. Environmental signals controlling expression of virulence determinants in bacteria. J Bacteriol. 1992 Jan;174(1):1–7. doi: 10.1128/jb.174.1.1-7.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. O'Toole P. W., O'Toole P., Stenberg L., Rissler M., Lindahl G. Two major classes in the M protein family in group A streptococci. Proc Natl Acad Sci U S A. 1992 Sep 15;89(18):8661–8665. doi: 10.1073/pnas.89.18.8661. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Okada N., Geist R. T., Caparon M. G. Positive transcriptional control of mry regulates virulence in the group A streptococcus. Mol Microbiol. 1993 Mar;7(6):893–903. doi: 10.1111/j.1365-2958.1993.tb01180.x. [DOI] [PubMed] [Google Scholar]
  32. Pancholi V., Fischetti V. A. A major surface protein on group A streptococci is a glyceraldehyde-3-phosphate-dehydrogenase with multiple binding activity. J Exp Med. 1992 Aug 1;176(2):415–426. doi: 10.1084/jem.176.2.415. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Perez-Casal J. F., Dillon H. F., Husmann L. K., Graham B., Scott J. R. Virulence of two Streptococcus pyogenes strains (types M1 and M3) associated with toxic-shock-like syndrome depends on an intact mry-like gene. Infect Immun. 1993 Dec;61(12):5426–5430. doi: 10.1128/iai.61.12.5426-5430.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Perez-Casal J., Caparon M. G., Scott J. R. Mry, a trans-acting positive regulator of the M protein gene of Streptococcus pyogenes with similarity to the receptor proteins of two-component regulatory systems. J Bacteriol. 1991 Apr;173(8):2617–2624. doi: 10.1128/jb.173.8.2617-2624.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Podbielski A., Hawlitzky J., Pack T. D., Flosdorff A., Boyle M. D. A group A streptococcal Enn protein potentially resulting from intergenomic recombination exhibits atypical immunoglobulin-binding characteristics. Mol Microbiol. 1994 Jun;12(5):725–736. doi: 10.1111/j.1365-2958.1994.tb01060.x. [DOI] [PubMed] [Google Scholar]
  36. Podbielski A., Krebs B., Kaufhold A. Genetic variability of the emm-related gene of the large vir regulon of group A streptococci: potential intra- and intergenomic recombination events. Mol Gen Genet. 1994 Jun 15;243(6):691–698. doi: 10.1007/BF00279579. [DOI] [PubMed] [Google Scholar]
  37. Podbielski A., Peterson J. A., Cleary P. Surface protein-CAT reporter fusions demonstrate differential gene expression in the vir regulon of Streptococcus pyogenes. Mol Microbiol. 1992 Aug;6(16):2253–2265. doi: 10.1111/j.1365-2958.1992.tb01401.x. [DOI] [PubMed] [Google Scholar]
  38. Podbielski A., Schönling J., Melzer B., Haase G. Different promoters of SHV-2 and SHV-2a beta-lactamase lead to diverse levels of cefotaxime resistance in their bacterial producers. J Gen Microbiol. 1991 Jul;137(7):1667–1675. doi: 10.1099/00221287-137-7-1667. [DOI] [PubMed] [Google Scholar]
  39. Podbielski A. Three different types of organization of the vir regulon in group A streptococci. Mol Gen Genet. 1993 Feb;237(1-2):287–300. doi: 10.1007/BF00282810. [DOI] [PubMed] [Google Scholar]
  40. Podbielski A. Ubiquitous occurrence of virR and scpA genes in group A streptococci. Med Microbiol Immunol. 1992;181(4):227–240. doi: 10.1007/BF00215768. [DOI] [PubMed] [Google Scholar]
  41. Raeder R., Boyle M. D. Association between expression of immunoglobulin G-binding proteins by group A streptococci and virulence in a mouse skin infection model. Infect Immun. 1993 Apr;61(4):1378–1384. doi: 10.1128/iai.61.4.1378-1384.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Raeder R., Boyle M. D. Association of type II immunoglobulin G-binding protein expression and survival of group A streptococci in human blood. Infect Immun. 1993 Sep;61(9):3696–3702. doi: 10.1128/iai.61.9.3696-3702.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Richardson J. P. Transcription termination. Crit Rev Biochem Mol Biol. 1993;28(1):1–30. doi: 10.3109/10409239309082571. [DOI] [PubMed] [Google Scholar]
  44. Sela S., Aviv A., Tovi A., Burstein I., Caparon M. G., Hanski E. Protein F: an adhesin of Streptococcus pyogenes binds fibronectin via two distinct domains. Mol Microbiol. 1993 Dec;10(5):1049–1055. doi: 10.1111/j.1365-2958.1993.tb00975.x. [DOI] [PubMed] [Google Scholar]
  45. Simpson W. J., Cleary P. P. Expression of M type 12 protein by a group A streptococcus exhibits phaselike variation: evidence for coregulation of colony opacity determinants and M protein. Infect Immun. 1987 Oct;55(10):2448–2455. doi: 10.1128/iai.55.10.2448-2455.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  46. Simpson W. J., LaPenta D., Chen C., Cleary P. P. Coregulation of type 12 M protein and streptococcal C5a peptidase genes in group A streptococci: evidence for a virulence regulon controlled by the virR locus. J Bacteriol. 1990 Feb;172(2):696–700. doi: 10.1128/jb.172.2.696-700.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  47. Stevens D. L. Invasive group A streptococcus infections. Clin Infect Dis. 1992 Jan;14(1):2–11. doi: 10.1093/clinids/14.1.2. [DOI] [PubMed] [Google Scholar]
  48. Tao L., LeBlanc D. J., Ferretti J. J. Novel streptococcal-integration shuttle vectors for gene cloning and inactivation. Gene. 1992 Oct 12;120(1):105–110. doi: 10.1016/0378-1119(92)90016-i. [DOI] [PubMed] [Google Scholar]
  49. Wannamaker L. W. Differences between streptococcal infections of the throat and of the skin (second of two parts). N Engl J Med. 1970 Jan 8;282(2):78–85. doi: 10.1056/NEJM197001082820206. [DOI] [PubMed] [Google Scholar]
  50. Zhou C., Yang Y., Jong A. Y. Mini-prep in ten minutes. Biotechniques. 1990 Feb;8(2):172–173. [PubMed] [Google Scholar]

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