Skip to main content
Infection and Immunity logoLink to Infection and Immunity
. 1996 Jun;64(6):2193–2200. doi: 10.1128/iai.64.6.2193-2200.1996

DNA sequencing and gene expression of the emm gene cluster in an M50 group A streptococcus strain virulent for mice.

D L Yung 1, S K Hollingshead 1
PMCID: PMC174055  PMID: 8675326

Abstract

The strain B514, an M serotype 50 strain, is capable of causing a natural upper respiratory infection leading to death in mice, as reported by Hook et al. in 1960 (E. W. Hook, R. R. Wagner, and R. C. Lancefield, Am. J. Hyg. 72:111-119, 1960). Thus, this strain was of interest for use in developing an animal model for group A streptococcal colonization and disease. The emm gene cluster for this strain was examined by PCR mapping and found to contain three emm family genes and cluster pattern 5. PCR-generated fragments corresponding to the SF4 (mrp50), SF2 (emmL50), and SF3 (enn50) genes were cloned and the entire gene cluster was sequenced. The gene cluster has greater than 97% DNA identity to previously sequenced regions of the gene cluster of the M2 strain T2/44/RB4 if two small divergent regions that encode the mature amino terminus of the SF-2 and SF-3 gene products are not included. If expressed, the genes encode proteins which bind human immunoglobulin G (Mrp50 and EmmL50) or immunoglobulin A (Enn50). However, in isolates taken directly after passage in mice, the surface proteins arising from these genes were barely detectable. The transcription of each gene in the B514 strain was investigated by Northern (RNA) hybridization, and mRNA transcripts were detected and quantitated relative to those of the recA gene, a housekeeping gene. Transcription of all three emm family genes was found to be over 30-fold attenuated relative to transcription of the same genes in strain T2/44/RB4. This suggests that the positive regulator, Mga, either is not expressed in this strain or has a different requirement for activation; it also suggests that the capsule may be sufficient to inhibit phagocytosis under these circumstances.

Full Text

The Full Text of this article is available as a PDF (518.5 KB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Berge A., Sjöbring U. PAM, a novel plasminogen-binding protein from Streptococcus pyogenes. J Biol Chem. 1993 Dec 5;268(34):25417–25424. [PubMed] [Google Scholar]
  2. 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]
  3. 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]
  4. Bessen D. E., Hollingshead S. K. Allelic polymorphism of emm loci provides evidence for horizontal gene spread in group A streptococci. Proc Natl Acad Sci U S A. 1994 Apr 12;91(8):3280–3284. doi: 10.1073/pnas.91.8.3280. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Bessen D. E., Hollingshead S. K. Horizontal transfer and mosaic-like emm gene structures in group A streptococci. Dev Biol Stand. 1995;85:169–173. [PubMed] [Google Scholar]
  6. Bessen D. E. Localization of immunoglobulin A-binding sites within M or M-like proteins of group A streptococci. Infect Immun. 1994 May;62(5):1968–1974. doi: 10.1128/iai.62.5.1968-1974.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. 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]
  8. 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]
  9. Dybvig K., Hollingshead S. K., Heath D. G., Clewell D. B., Sun F., Woodard A. Degenerate oligonucleotide primers for enzymatic amplification of recA sequences from gram-positive bacteria and mycoplasmas. J Bacteriol. 1992 Apr;174(8):2729–2732. doi: 10.1128/jb.174.8.2729-2732.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Frick I. M., Akesson P., Cooney J., Sjöbring U., Schmidt K. H., Gomi H., Hattori S., Tagawa C., Kishimoto F., Björck L. Protein H--a surface protein of Streptococcus pyogenes with separate binding sites for IgG and albumin. Mol Microbiol. 1994 Apr;12(1):143–151. doi: 10.1111/j.1365-2958.1994.tb01003.x. [DOI] [PubMed] [Google Scholar]
  11. HOOK E. W., WAGNER R. R., LANCEFIELD R. C. An epizootic in Swiss mice caused by a group A Streptococcus, newly designated type 50. Am J Hyg. 1960 Jul;72:111–119. doi: 10.1093/oxfordjournals.aje.a120127. [DOI] [PubMed] [Google Scholar]
  12. 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]
  13. 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]
  14. Hedén L. O., Lindahl G. Conserved and variable regions in protein Arp, the IgA receptor of Streptococcus pyogenes. J Gen Microbiol. 1993 Sep;139(9):2067–2074. doi: 10.1099/00221287-139-9-2067. [DOI] [PubMed] [Google Scholar]
  15. Hollingshead S. K., Arnold J., Readdy T. L., Bessen D. E. Molecular evolution of a multigene family in group A streptococci. Mol Biol Evol. 1994 Mar;11(2):208–219. doi: 10.1093/oxfordjournals.molbev.a040103. [DOI] [PubMed] [Google Scholar]
  16. Hollingshead S. K., Bessen D. E. Evolution of the emm gene family: virulence gene clusters in group A streptococci. Dev Biol Stand. 1995;85:163–168. [PubMed] [Google Scholar]
  17. Hollingshead S. K., Fischetti V. A., Scott J. R. Complete nucleotide sequence of type 6 M protein of the group A Streptococcus. Repetitive structure and membrane anchor. J Biol Chem. 1986 Feb 5;261(4):1677–1686. [PubMed] [Google Scholar]
  18. Hollingshead S. K., Fischetti V. A., Scott J. R. Size variation in group A streptococcal M protein is generated by homologous recombination between intragenic repeats. Mol Gen Genet. 1987 May;207(2-3):196–203. doi: 10.1007/BF00331578. [DOI] [PubMed] [Google Scholar]
  19. 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]
  20. Horstmann R. D., Sievertsen H. J., Knobloch J., Fischetti V. A. Antiphagocytic activity of streptococcal M protein: selective binding of complement control protein factor H. Proc Natl Acad Sci U S A. 1988 Mar;85(5):1657–1661. doi: 10.1073/pnas.85.5.1657. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Jones K. F., Fischetti V. A. The importance of the location of antibody binding on the M6 protein for opsonization and phagocytosis of group A M6 streptococci. J Exp Med. 1988 Mar 1;167(3):1114–1123. doi: 10.1084/jem.167.3.1114. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Kraus W., Haanes-Fritz E., Cleary P. P., Seyer J. M., Dale J. B., Beachey E. H. Sequence and type-specific immunogenicity of the amino-terminal region of type 1 streptococcal M protein. J Immunol. 1987 Nov 1;139(9):3084–3090. [PubMed] [Google Scholar]
  23. Kurl D. N., Stjernquist-Desatnik A., Schalén C., Christensen P. Induction of local immunity to group A streptococci type M50 in mice by non-type-specific mechanisms. Acta Pathol Microbiol Immunol Scand B. 1985 Dec;93(6):401–405. doi: 10.1111/j.1699-0463.1985.tb02909.x. [DOI] [PubMed] [Google Scholar]
  24. Lindahl G. Cell surface proteins of a group A streptococcus type M4: the IgA receptor and a receptor related to M proteins are coded for by closely linked genes. Mol Gen Genet. 1989 Apr;216(2-3):372–379. doi: 10.1007/BF00334378. [DOI] [PubMed] [Google Scholar]
  25. McIver K. S., Heath A. S., Green B. D., Scott J. R. Specific binding of the activator Mga to promoter sequences of the emm and scpA genes in the group A streptococcus. J Bacteriol. 1995 Nov;177(22):6619–6624. doi: 10.1128/jb.177.22.6619-6624.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Musser J. M., Kapur V., Szeto J., Pan X., Swanson D. S., Martin D. R. Genetic diversity and relationships among Streptococcus pyogenes strains expressing serotype M1 protein: recent intercontinental spread of a subclone causing episodes of invasive disease. Infect Immun. 1995 Mar;63(3):994–1003. doi: 10.1128/iai.63.3.994-1003.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. 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]
  28. 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]
  29. Phillips G. N., Jr, Flicker P. F., Cohen C., Manjula B. N., Fischetti V. A. Streptococcal M protein: alpha-helical coiled-coil structure and arrangement on the cell surface. Proc Natl Acad Sci U S A. 1981 Aug;78(8):4689–4693. doi: 10.1073/pnas.78.8.4689. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Podbielski A., Flosdorff A., Weber-Heynemann J. The group A streptococcal virR49 gene controls expression of four structural vir regulon genes. Infect Immun. 1995 Jan;63(1):9–20. doi: 10.1128/iai.63.1.9-20.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. 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]
  32. Raeder R., Otten R. A., Chamberlin L., Boyle M. D. Functional and serological analysis of type II immunoglobulin G-binding proteins expressed by pathogenic group A streptococci. J Clin Microbiol. 1992 Dec;30(12):3074–3081. doi: 10.1128/jcm.30.12.3074-3081.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Rakonjac J. V., Robbins J. C., Fischetti V. A. DNA sequence of the serum opacity factor of group A streptococci: identification of a fibronectin-binding repeat domain. Infect Immun. 1995 Feb;63(2):622–631. doi: 10.1128/iai.63.2.622-631.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Retnoningrum D. S., Cleary P. P. M12 protein from Streptococcus pyogenes is a receptor for immunoglobulin G3 and human albumin. Infect Immun. 1994 Jun;62(6):2387–2394. doi: 10.1128/iai.62.6.2387-2394.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Robbins J. C., Spanier J. G., Jones S. J., Simpson W. J., Cleary P. P. Streptococcus pyogenes type 12 M protein gene regulation by upstream sequences. J Bacteriol. 1987 Dec;169(12):5633–5640. doi: 10.1128/jb.169.12.5633-5640.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Rotta J., Krause R. M., Lancefield R. C., Everly W., Lackland H. New approaches for the laboratory recognition of M types of group A streptococci. J Exp Med. 1971 Nov 1;134(5):1298–1315. doi: 10.1084/jem.134.5.1298. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Schalén C. Prevalence of IgA receptors in clinical isolates of Streptococcus pyogenes and Streptococcus agalactiae: serologic distinction between the receptors by blocking antibodies. FEMS Immunol Med Microbiol. 1993 Jun;7(1):39–45. doi: 10.1111/j.1574-695X.1993.tb00379.x. [DOI] [PubMed] [Google Scholar]
  39. Scott J. R., Cleary P., Caparon M. G., Kehoe M., Heden L., Musser J. M., Hollingshead S., Podbielski A. New name for the positive regulator of the M protein of group A streptococcus. Mol Microbiol. 1995 Aug;17(4):799–799. doi: 10.1111/j.1365-2958.1995.mmi_17040799.x. [DOI] [PubMed] [Google Scholar]
  40. Stenberg L., O'Toole P., Lindahl G. Many group A streptococcal strains express two different immunoglobulin-binding proteins, encoded by closely linked genes: characterization of the proteins expressed by four strains of different M-type. Mol Microbiol. 1992 May;6(9):1185–1194. doi: 10.1111/j.1365-2958.1992.tb01557.x. [DOI] [PubMed] [Google Scholar]
  41. Stjernquist-Desatnik A., Kurl D. N., Schalén C., Christensen P. Protective effect of heterologous gram-positive vaccine against lethal upper respiratory tract infection with type M50 group A streptococci in mice. Vaccine. 1990 Apr;8(2):150–152. doi: 10.1016/0264-410x(90)90138-c. [DOI] [PubMed] [Google Scholar]
  42. Thern A., Stenberg L., Dahlbäck B., Lindahl G. Ig-binding surface proteins of Streptococcus pyogenes also bind human C4b-binding protein (C4BP), a regulatory component of the complement system. J Immunol. 1995 Jan 1;154(1):375–386. [PubMed] [Google Scholar]
  43. Wessels M. R., Bronze M. S. Critical role of the group A streptococcal capsule in pharyngeal colonization and infection in mice. Proc Natl Acad Sci U S A. 1994 Dec 6;91(25):12238–12242. doi: 10.1073/pnas.91.25.12238. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. Whatmore A. M., Kehoe M. A. Horizontal gene transfer in the evolution of group A streptococcal emm-like genes: gene mosaics and variation in Vir regulons. Mol Microbiol. 1994 Jan;11(2):363–374. doi: 10.1111/j.1365-2958.1994.tb00316.x. [DOI] [PubMed] [Google Scholar]
  45. Whitnack E., Dale J. B., Beachey E. H. Common protective antigens of group A streptococcal M proteins masked by fibrinogen. J Exp Med. 1984 Apr 1;159(4):1201–1212. doi: 10.1084/jem.159.4.1201. [DOI] [PMC free article] [PubMed] [Google Scholar]
  46. Wildfeuer A., Heymer B., Schachenmayr W., Haferkamp O. Morphological and immunological characteristics of Streptococcus pyogenes, group A, type 50. Med Microbiol Immunol. 1975 Jul 2;161(3):193–201. doi: 10.1007/BF02121010. [DOI] [PubMed] [Google Scholar]
  47. Wildfeuer A., Heymer B., Schäfer H., Haferkamp O. The cervical lymph nodes in Streptococcus pyogenes, group A, type 50, infection in mice. Med Microbiol Immunol. 1978 Jul 4;165(2):129–137. doi: 10.1007/BF02122748. [DOI] [PubMed] [Google Scholar]

Articles from Infection and Immunity are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES