Skip to main content
NCBI home page
Infection and Immunity logoLink to Infection and Immunity
. 1987 Oct;55(10):2448–2455. doi: 10.1128/iai.55.10.2448-2455.1987

Expression of M type 12 protein by a group A streptococcus exhibits phaselike variation: evidence for coregulation of colony opacity determinants and M protein.

W J Simpson 1, P P Cleary 1
PMCID: PMC260728  PMID: 2443452

Abstract

Three major categories of colony opacity were observed for natural variants of the M type 12 (M12) group A streptococcus strain CS24. Colony opacity variants that switched between two alternative categories at significantly high frequencies were identified and are referred to as switching between more opaque (Op+) and less opaque (Op-) phenotypes. Twenty lineages of such variants were derived for analysis and were assessed for resistance to phagocytosis, acid-extractable M12 antigen, and M12 mRNA, criteria which define the M protein-positive phenotype (M+). Transition from the M+ to the M protein-negative phenotype (M-) correlated with a change from Op+ to Op-. Reversion to the Op+ phenotype was accompanied by reversion to the M+ state in all variants except one and occurred at a higher frequency than the forward M+ to M- switch. These data demonstrate the existence of M12 protein phaselike switching in the group A streptococcus strain CS24. The discovery of an Op+ M- revertant confirmed that colony opacity and M protein can be expressed independently and are distinct gene products. We suggest that coregulation of colony opacity and M protein expression accounts for their association among descendents of strain CS24. Southern blot hybridization analyses of digested genomic DNA from 27 M- variants and 15 M+ revertants were performed with DNA probes containing M12 protein and adjacent upstream sequences. DNA deletions were identified only in two stable M- variants, approximately 1.3 and 1.4 kilobases upstream from the M12 gene, respectively, whereas all unstable M- variants lacked detectable rearrangements. This suggests that deletions within or adjacent to the structural gene are unlikely to be responsible for the reversible switch in M protein expression. However, the association with the stable M- phenotype and the location of these deletions, as well as two other deletions, approximately 0.5 kilobase upstream from the M12 promoter in two previously described variants of strain CS24 suggests that a second gene product is required for full expression of M12 protein synthesis in this strain.

Full text

PDF
2448

Images in this article

2450Image
on p.2450
2452Image
on p.2452
2452Image
on p.2452

Selected References

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

  1. Beachey E. H., Stollerman G. H. Toxic effects of streptococcal M protein on platelets and polymorphonuclear leukocytes in human blood. J Exp Med. 1971 Aug 1;134(2):351–365. doi: 10.1084/jem.134.2.351. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bisno A. L. Alternate complement pathway activation by group A streptococci: role of M-protein. Infect Immun. 1979 Dec;26(3):1172–1176. doi: 10.1128/iai.26.3.1172-1176.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Cleary P. P., Johnson Z., Wannamaker L. Genetic instability of M protein and serum opacity factor of group A streptocci: evidence suggesting extrachromosomal control. Infect Immun. 1975 Jul;12(1):109–118. doi: 10.1128/iai.12.1.109-118.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Eisenstein B. I. Phase variation of type 1 fimbriae in Escherichia coli is under transcriptional control. Science. 1981 Oct 16;214(4518):337–339. doi: 10.1126/science.6116279. [DOI] [PubMed] [Google Scholar]
  5. Goodman H. M., MacDonald R. J. Cloning of hormone genes from a mixture of cDNA molecules. Methods Enzymol. 1979;68:75–90. doi: 10.1016/0076-6879(79)68007-2. [DOI] [PubMed] [Google Scholar]
  6. Hagblom P., Segal E., Billyard E., So M. Intragenic recombination leads to pilus antigenic variation in Neisseria gonorrhoeae. Nature. 1985 May 9;315(6015):156–158. doi: 10.1038/315156a0. [DOI] [PubMed] [Google Scholar]
  7. Hu N., Messing J. The making of strand-specific M13 probes. Gene. 1982 Mar;17(3):271–277. doi: 10.1016/0378-1119(82)90143-3. [DOI] [PubMed] [Google Scholar]
  8. 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]
  9. Lancefield R. C. THE ANTIGENIC COMPLEX OF STREPTOCOCCUS HAEMOLYTICUS : I. DEMONSTRATION OF A TYPE-SPECIFIC SUBSTANCE IN EXTRACTS OF STREPTOCOCCUS HAEMOLYTICUS. J Exp Med. 1928 Jan 1;47(1):91–103. doi: 10.1084/jem.47.1.91. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Manjula B. N., Schmidt M. L., Fischetti V. A. Unimpaired function of human phagocytes in the presence of phagocytosis-resistant group A streptococci. Infect Immun. 1985 Dec;50(3):610–613. doi: 10.1128/iai.50.3.610-613.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Maxted W. R., Valkenburg H. A. Variation in the M-antigen of group-A streptococci. J Med Microbiol. 1969 Aug;2(3):199–210. doi: 10.1099/00222615-2-3-199. [DOI] [PubMed] [Google Scholar]
  12. McCarty M. The nature of the opaque colony variation in group A streptococci. J Hyg (Lond) 1966 Jun;64(2):185–190. doi: 10.1017/s0022172400040444. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Minges C. G., Titus J. A., Strohl W. R. Plasmid DNA in colorless filamentous gliding bacteria. Arch Microbiol. 1983 Jan;134(1):38–44. doi: 10.1007/BF00429404. [DOI] [PubMed] [Google Scholar]
  14. ROTHBARD S., WATSON R. F. Variation occurring in group A streptococci during human infection; progressive loss of M substance correlated with increasing susceptibility to bacteriostasis. J Exp Med. 1948 Jun 1;87(6):521–533. doi: 10.1084/jem.87.6.521. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Recsei P., Kreiswirth B., O'Reilly M., Schlievert P., Gruss A., Novick R. P. Regulation of exoprotein gene expression in Staphylococcus aureus by agar. Mol Gen Genet. 1986 Jan;202(1):58–61. doi: 10.1007/BF00330517. [DOI] [PubMed] [Google Scholar]
  16. Skjold S. A., Wannamaker L. W. Surface proteins in the transduction of groups A and G streptococci. J Med Microbiol. 1986 Feb;21(1):69–74. doi: 10.1099/00222615-21-1-69. [DOI] [PubMed] [Google Scholar]
  17. Spanier J. G., Cleary P. P. A DNA substitution in the group A streptococcal bacteriophage SP24. Virology. 1983 Oct 30;130(2):514–522. doi: 10.1016/0042-6822(83)90104-6. [DOI] [PubMed] [Google Scholar]
  18. Spanier J. G., Cleary P. P. Bacteriophage control of antiphagocytic determinants in group A streptococci. J Exp Med. 1980 Nov 1;152(5):1393–1406. doi: 10.1084/jem.152.5.1393. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Spanier J. G., Cleary P. P. Integration of bacteriophage SP24 into the chromosome of group A streptococci. J Bacteriol. 1985 Nov;164(2):600–604. doi: 10.1128/jb.164.2.600-604.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Spanier J. G., Jones S. J., Cleary P. Small DNA deletions creating avirulence in Streptococcus pyogenes. Science. 1984 Aug 31;225(4665):935–938. doi: 10.1126/science.6089334. [DOI] [PubMed] [Google Scholar]
  21. Sparling P. F., Cannon J. G., So M. Phase and antigenic variation of pili and outer membrane protein II of Neisseria gonorrhoeae. J Infect Dis. 1986 Feb;153(2):196–201. doi: 10.1093/infdis/153.2.196. [DOI] [PubMed] [Google Scholar]
  22. Swanson J., McCarty M. Electron microscopic studies on opaque colony variants of group A streptococci. J Bacteriol. 1969 Oct;100(1):505–511. doi: 10.1128/jb.100.1.505-511.1969. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Thomas P. S. Hybridization of denatured RNA and small DNA fragments transferred to nitrocellulose. Proc Natl Acad Sci U S A. 1980 Sep;77(9):5201–5205. doi: 10.1073/pnas.77.9.5201. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Vieira J., Messing J. The pUC plasmids, an M13mp7-derived system for insertion mutagenesis and sequencing with synthetic universal primers. Gene. 1982 Oct;19(3):259–268. doi: 10.1016/0378-1119(82)90015-4. [DOI] [PubMed] [Google Scholar]
  25. WILSON A. T. The relative importance of the capsule and the M-antigen in determining colony form of group A streptococci. J Exp Med. 1959 Mar 1;109(3):257–270. doi: 10.1084/jem.109.3.257. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Zigmond S. H., Hirsch J. G. Effects of cytochalasin B on polymorphonuclear leucocyte locomotion, phagocytosis and glycolysis. Exp Cell Res. 1972 Aug;73(2):383–393. doi: 10.1016/0014-4827(72)90062-6. [DOI] [PubMed] [Google Scholar]

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

RESOURCES