Skip to main content
Infection and Immunity logoLink to Infection and Immunity
. 1994 Apr;62(4):1336–1347. doi: 10.1128/iai.62.4.1336-1347.1994

Analysis of genes encoding two unique type IIa immunoglobulin G-binding proteins expressed by a single group A streptococcal isolate.

M D Boyle 1, J Hawlitzky 1, R Raeder 1, A Podbielski 1
PMCID: PMC186283  PMID: 8132341

Abstract

An emm-like gene (emmL) and a fcrA gene from group A streptococcal strain 64/14 (emmL64/14 and fcrA64/14) were amplified by PCR and force cloned into the heat-inducible expression vector pJLA 602. The emmL gene encoded a recombinant protein that bound human IgG1, IgG2, and IgG4 in a nonimmune fashion. This is the reactivity profile of a type IIa IgG-binding protein. The emmL64/14 gene product was antigenically similar to the previously identified high-molecular-weight type IIa IgG-binding protein of strain 64/14 and had an N-terminal sequence identical to that of the wild-type protein. The fcrA gene also encoded a recombinant protein with type IIa functional activity. This protein was similar to the lower-molecular-weight type IIa IgG-binding protein previously isolated from strain 64/14 and was antigenically distinct from the higher-molecular-weight type IIa protein encoded by the emmL64/14 gene. The sequences for both genes including the intervening regions are presented. The emmL gene demonstrates significant homology to other class I emm and emmL genes expressed by opacity factor-negative group A streptococcal isolates. The fcrA gene was found to be homologous to other fcrA genes normally present in opacity factor-positive group A isolates. The sequence upstream of the fcrA gene and the intervening sequence between the end of the fcrA gene and the start of the emmL gene were similar to those reported for other fcrA genes.

Full text

PDF
1338

Images in this article

Selected References

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

  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. Bessen D., Jones K. F., Fischetti V. A. Evidence for two distinct classes of streptococcal M protein and their relationship to rheumatic fever. J Exp Med. 1989 Jan 1;169(1):269–283. doi: 10.1084/jem.169.1.269. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Burova L. A., Koroleva I. V., Ogurtzov R. P., Murashov S. V., Svensson M. L., Schalén C. Role of streptococcal IgG Fc receptor in tissue deposition of IgG in rabbits immunized with Streptococcus pyogenes. APMIS. 1992 Jun;100(6):567–574. doi: 10.1111/j.1699-0463.1992.tb00912.x. [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., 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]
  8. Crowe J. S., Cooper H. J., Smith M. A., Sims M. J., Parker D., Gewert D. Improved cloning efficiency of polymerase chain reaction (PCR) products after proteinase K digestion. Nucleic Acids Res. 1991 Jan 11;19(1):184–184. doi: 10.1093/nar/19.1.184. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. 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]
  10. Fischetti V. A. Streptococcal M protein: molecular design and biological behavior. Clin Microbiol Rev. 1989 Jul;2(3):285–314. doi: 10.1128/cmr.2.3.285. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Frithz E., Hedén L. O., Lindahl G. Extensive sequence homology between IgA receptor and M proteins in Streptococcus pyogenes. Mol Microbiol. 1989 Aug;3(8):1111–1119. doi: 10.1111/j.1365-2958.1989.tb00261.x. [DOI] [PubMed] [Google Scholar]
  12. Gomi H., Hozumi T., Hattori S., Tagawa C., Kishimoto F., Björck L. The gene sequence and some properties of protein H. A novel IgG-binding protein. J Immunol. 1990 May 15;144(10):4046–4052. [PubMed] [Google Scholar]
  13. 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]
  14. 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]
  15. 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]
  16. Heath D. G., Cleary P. P. Cloning and expression of the gene for an immunoglobulin G Fc receptor protein from a group A streptococcus. Infect Immun. 1987 May;55(5):1233–1238. doi: 10.1128/iai.55.5.1233-1238.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. 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]
  18. 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]
  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. 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]
  21. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  22. Martin B., Alloing G., Boucraut C., Claverys J. P. The difficulty of cloning Streptococcus pneumoniae mal and ami loci in Escherichia coli: toxicity of malX and amiA gene products. Gene. 1989 Aug 15;80(2):227–238. doi: 10.1016/0378-1119(89)90287-4. [DOI] [PubMed] [Google Scholar]
  23. 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]
  24. 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]
  25. Otten R. A., Raeder R., Heath D. G., Lottenberg R., Cleary P. P., Boyle M. D. Identification of two type IIa IgG-binding proteins expressed by a single group A streptococcus. J Immunol. 1992 May 15;148(10):3174–3182. [PubMed] [Google Scholar]
  26. 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]
  27. Podbielski A., Melzer B., Lütticken R. Application of the polymerase chain reaction to study the M protein(-like) gene family in beta-hemolytic streptococci. Med Microbiol Immunol. 1991;180(4):213–227. doi: 10.1007/BF00215250. [DOI] [PubMed] [Google Scholar]
  28. 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]
  29. 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]
  30. 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]
  31. Raeder R., Faulmann E. L., Boyle M. D. Evidence for functional heterogeneity in IgG Fc-binding proteins associated with group A streptococci. J Immunol. 1991 Feb 15;146(4):1247–1253. [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. Reis K. J., Yarnall M., Ayoub E. M., Boyle M. D. Effect of mouse passage on Fc receptor expression by group A streptococci. Scand J Immunol. 1984 Nov;20(5):433–439. doi: 10.1111/j.1365-3083.1984.tb01022.x. [DOI] [PubMed] [Google Scholar]
  34. Retnoningrum D. S., Podbielski A., Cleary P. P. Type M12 protein from Streptococcus pyogenes is a receptor for IgG3. J Immunol. 1993 Mar 15;150(6):2332–2340. [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. Saiki R. K., Gelfand D. H., Stoffel S., Scharf S. J., Higuchi R., Horn G. T., Mullis K. B., Erlich H. A. Primer-directed enzymatic amplification of DNA with a thermostable DNA polymerase. Science. 1988 Jan 29;239(4839):487–491. doi: 10.1126/science.2448875. [DOI] [PubMed] [Google Scholar]
  37. Schmidt K. H., Wadström T. A secreted receptor related to M1 protein of Streptococcus pyogenes binds to fibrinogen, IgG, and albumin. Zentralbl Bakteriol. 1990 Jun;273(2):216–228. doi: 10.1016/s0934-8840(11)80252-5. [DOI] [PubMed] [Google Scholar]
  38. 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]
  39. 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]
  40. Towbin H., Staehelin T., Gordon J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4350–4354. doi: 10.1073/pnas.76.9.4350. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Whitehouse E. H., Spears T. A simple method for removing oil from cycle sequencing reactions. Biotechniques. 1991 Nov;11(5):616–618. [PubMed] [Google Scholar]
  42. Yarnall M., Boyle M. D. Isolation and characterization of type IIa and type IIb Fc receptors from a group A streptococcus. Scand J Immunol. 1986 Nov;24(5):549–557. doi: 10.1111/j.1365-3083.1986.tb02170.x. [DOI] [PubMed] [Google Scholar]
  43. Zhou C., Yang Y., Jong A. Y. Mini-prep in ten minutes. Biotechniques. 1990 Feb;8(2):172–173. [PubMed] [Google Scholar]

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

RESOURCES