Skip to main content
PMC home page
Infection and Immunity logoLink to Infection and Immunity
. 1996 Oct;64(10):3967–3974. doi: 10.1128/iai.64.10.3967-3974.1996

Identification, cloning, and sequencing of the immunoglobulin A1 protease gene of Streptococcus pneumoniae.

J H Wani 1, J V Gilbert 1, A G Plaut 1, J N Weiser 1
PMCID: PMC174324  PMID: 8926056

Abstract

The pneumococcus expresses a protease that hydrolyzes human immunoglobulin A1 (IgA1). A gene for IgA1 protease was identified from a plasmid library of pneumococcal DNA because of the effect of its overexpression on the colony morphology of Streptococcus pneumoniae. The deduced 1,964-amino-acid sequence is highly homologous to that of the IgA1 protease from Streptococcus sanguis. The similarity to the S. sanguis enzyme and the presence of a putative zinc-binding site suggest that the pneumococcal enzyme is a metalloprotease. The two streptococcal sequences differ in a hydrophilic region with 10 tandem repeats of a 20-mer in S. sanguis, which is replaced by a similar but less repetitive sequence in S. pneumoniae. Antiserum reactive with the pneumococcal IgA1 protease was used to demonstrate that the majority of the protein is cell associated. The expression and function of this gene were confirmed by insertional mutagenesis. Interruption of the chromosomal gene resulted in loss of expression of an approximately 200-kDa protein and complete elimination of detectable IgA1 protease activity.

Full Text

The Full Text of this article is available as a PDF (1.2 MB).

Selected References

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

  1. Achen M. G., Davidson B. E., Hillier A. J. Construction of plasmid vectors for the detection of streptococcal promoters. Gene. 1986;45(1):45–49. doi: 10.1016/0378-1119(86)90130-7. [DOI] [PubMed] [Google Scholar]
  2. Altschul S. F., Gish W., Miller W., Myers E. W., Lipman D. J. Basic local alignment search tool. J Mol Biol. 1990 Oct 5;215(3):403–410. doi: 10.1016/S0022-2836(05)80360-2. [DOI] [PubMed] [Google Scholar]
  3. Bricker J., Mulks M. H., Plaut A. G., Moxon E. R., Wright A. IgA1 proteases of Haemophilus influenzae: cloning and characterization in Escherichia coli K-12. Proc Natl Acad Sci U S A. 1983 May;80(9):2681–2685. doi: 10.1073/pnas.80.9.2681. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Devereux J., Haeberli P., Smithies O. A comprehensive set of sequence analysis programs for the VAX. Nucleic Acids Res. 1984 Jan 11;12(1 Pt 1):387–395. doi: 10.1093/nar/12.1part1.387. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Fischetti V. A., Pancholi V., Schneewind O. Conservation of a hexapeptide sequence in the anchor region of surface proteins from gram-positive cocci. Mol Microbiol. 1990 Sep;4(9):1603–1605. doi: 10.1111/j.1365-2958.1990.tb02072.x. [DOI] [PubMed] [Google Scholar]
  6. Gilbert J. V., Plaut A. G., Fishman Y., Wright A. Cloning of the gene encoding streptococcal immunoglobulin A protease and its expression in Escherichia coli. Infect Immun. 1988 Aug;56(8):1961–1966. doi: 10.1128/iai.56.8.1961-1966.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Gilbert J. V., Plaut A. G., Wright A. Analysis of the immunoglobulin A protease gene of Streptococcus sanguis. Infect Immun. 1991 Jan;59(1):7–17. doi: 10.1128/iai.59.1.7-17.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. 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]
  9. Kett K., Brandtzaeg P., Radl J., Haaijman J. J. Different subclass distribution of IgA-producing cells in human lymphoid organs and various secretory tissues. J Immunol. 1986 May 15;136(10):3631–3635. [PubMed] [Google Scholar]
  10. Kilian M., Mestecky J., Schrohenloher R. E. Pathogenic species of the genus Haemophilus and Streptococcus pneumoniae produce immunoglobulin A1 protease. Infect Immun. 1979 Oct;26(1):143–149. doi: 10.1128/iai.26.1.143-149.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Koomey J. M., Falkow S. Nucleotide sequence homology between the immunoglobulin A1 protease genes of Neisseria gonorrhoeae, Neisseria meningitidis, and Haemophilus influenzae. Infect Immun. 1984 Jan;43(1):101–107. doi: 10.1128/iai.43.1.101-107.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Kyte J., Doolittle R. F. A simple method for displaying the hydropathic character of a protein. J Mol Biol. 1982 May 5;157(1):105–132. doi: 10.1016/0022-2836(82)90515-0. [DOI] [PubMed] [Google Scholar]
  13. LACKS S., HOTCHKISS R. D. A study of the genetic material determining an enzyme in Pneumococcus. Biochim Biophys Acta. 1960 Apr 22;39:508–518. doi: 10.1016/0006-3002(60)90205-5. [DOI] [PubMed] [Google Scholar]
  14. Labib R. S., Calvanico N. J., Tomasi T. B., Jr Studies on extracellular proteases of Streptococcus sanguis. Purification and characterization of a human IgA1 specific protease. Biochim Biophys Acta. 1978 Oct 12;526(2):547–559. doi: 10.1016/0005-2744(78)90145-6. [DOI] [PubMed] [Google Scholar]
  15. Lomholt H. Evidence of recombination and an antigenically diverse immunoglobulin A1 protease among strains of Streptococcus pneumoniae. Infect Immun. 1995 Nov;63(11):4238–4243. doi: 10.1128/iai.63.11.4238-4243.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Male C. J. Immunoglobulin A1 protease production by Haemophilus influenzae and Streptococcus pneumoniae. Infect Immun. 1979 Oct;26(1):254–261. doi: 10.1128/iai.26.1.254-261.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Monod M., Denoya C., Dubnau D. Sequence and properties of pIM13, a macrolide-lincosamide-streptogramin B resistance plasmid from Bacillus subtilis. J Bacteriol. 1986 Jul;167(1):138–147. doi: 10.1128/jb.167.1.138-147.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Mortensen S. B., Kilian M. Purification and characterization of an immunoglobulin A1 protease from Bacteroides melaninogenicus. Infect Immun. 1984 Sep;45(3):550–557. doi: 10.1128/iai.45.3.550-557.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Moxon E. R., Deich R. A., Connelly C. Cloning of chromosomal DNA from Haemophilus influenzae. Its use for studying the expression of type b capsule and virulence. J Clin Invest. 1984 Feb;73(2):298–306. doi: 10.1172/JCI111214. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Navarre W. W., Schneewind O. Proteolytic cleavage and cell wall anchoring at the LPXTG motif of surface proteins in gram-positive bacteria. Mol Microbiol. 1994 Oct;14(1):115–121. doi: 10.1111/j.1365-2958.1994.tb01271.x. [DOI] [PubMed] [Google Scholar]
  21. Plaut A. G., Gilbert J. V., Artenstein M. S., Capra J. D. Neisseria gonorrhoeae and neisseria meningitidis: extracellular enzyme cleaves human immunoglobulin A. Science. 1975 Dec 12;190(4219):1103–1105. doi: 10.1126/science.810892. [DOI] [PubMed] [Google Scholar]
  22. Plaut A. G. The IgA1 proteases of pathogenic bacteria. Annu Rev Microbiol. 1983;37:603–622. doi: 10.1146/annurev.mi.37.100183.003131. [DOI] [PubMed] [Google Scholar]
  23. Pohlner J., Halter R., Beyreuther K., Meyer T. F. Gene structure and extracellular secretion of Neisseria gonorrhoeae IgA protease. 1987 Jan 29-Feb 4Nature. 325(6103):458–462. doi: 10.1038/325458a0. [DOI] [PubMed] [Google Scholar]
  24. Poulsen K., Brandt J., Hjorth J. P., Thøgersen H. C., Kilian M. Cloning and sequencing of the immunoglobulin A1 protease gene (iga) of Haemophilus influenzae serotype b. Infect Immun. 1989 Oct;57(10):3097–3105. doi: 10.1128/iai.57.10.3097-3105.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Qiu J., Brackee G. P., Plaut A. G. Analysis of the specificity of bacterial immunoglobulin A (IgA) proteases by a comparative study of ape serum IgAs as substrates. Infect Immun. 1996 Mar;64(3):933–937. doi: 10.1128/iai.64.3.933-937.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Saluja S. K., Weiser J. N. The genetic basis of colony opacity in Streptococcus pneumoniae: evidence for the effect of box elements on the frequency of phenotypic variation. Mol Microbiol. 1995 Apr;16(2):215–227. doi: 10.1111/j.1365-2958.1995.tb02294.x. [DOI] [PubMed] [Google Scholar]
  27. 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]
  28. Saraste M., Sibbald P. R., Wittinghofer A. The P-loop--a common motif in ATP- and GTP-binding proteins. Trends Biochem Sci. 1990 Nov;15(11):430–434. doi: 10.1016/0968-0004(90)90281-f. [DOI] [PubMed] [Google Scholar]
  29. Schneewind O., Fowler A., Faull K. F. Structure of the cell wall anchor of surface proteins in Staphylococcus aureus. Science. 1995 Apr 7;268(5207):103–106. doi: 10.1126/science.7701329. [DOI] [PubMed] [Google Scholar]
  30. Schneewind O., Model P., Fischetti V. A. Sorting of protein A to the staphylococcal cell wall. Cell. 1992 Jul 24;70(2):267–281. doi: 10.1016/0092-8674(92)90101-h. [DOI] [PubMed] [Google Scholar]
  31. Shine J., Dalgarno L. The 3'-terminal sequence of Escherichia coli 16S ribosomal RNA: complementarity to nonsense triplets and ribosome binding sites. Proc Natl Acad Sci U S A. 1974 Apr;71(4):1342–1346. doi: 10.1073/pnas.71.4.1342. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Takeshita S., Sato M., Toba M., Masahashi W., Hashimoto-Gotoh T. High-copy-number and low-copy-number plasmid vectors for lacZ alpha-complementation and chloramphenicol- or kanamycin-resistance selection. Gene. 1987;61(1):63–74. doi: 10.1016/0378-1119(87)90365-9. [DOI] [PubMed] [Google Scholar]
  33. Tiraby J. G., Fox M. S. Marker discrimination in transformation and mutation of pneumococcus. Proc Natl Acad Sci U S A. 1973 Dec;70(12):3541–3545. doi: 10.1073/pnas.70.12.3541. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. 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]
  35. Tuomanen E. I., Austrian R., Masure H. R. Pathogenesis of pneumococcal infection. N Engl J Med. 1995 May 11;332(19):1280–1284. doi: 10.1056/NEJM199505113321907. [DOI] [PubMed] [Google Scholar]
  36. Vallee B. L., Auld D. S. Active-site zinc ligands and activated H2O of zinc enzymes. Proc Natl Acad Sci U S A. 1990 Jan;87(1):220–224. doi: 10.1073/pnas.87.1.220. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Vijayakumar M. N., Ayalew S. Nucleotide sequence analysis of the termini and chromosomal locus involved in site-specific integration of the streptococcal conjugative transposon Tn5252. J Bacteriol. 1993 May;175(9):2713–2719. doi: 10.1128/jb.175.9.2713-2719.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Weiser J. N., Austrian R., Sreenivasan P. K., Masure H. R. Phase variation in pneumococcal opacity: relationship between colonial morphology and nasopharyngeal colonization. Infect Immun. 1994 Jun;62(6):2582–2589. doi: 10.1128/iai.62.6.2582-2589.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Weiser J. N., Chong S. T., Greenberg D., Fong W. Identification and characterization of a cell envelope protein of Haemophilus influenzae contributing to phase variation in colony opacity and nasopharyngeal colonization. Mol Microbiol. 1995 Aug;17(3):555–564. doi: 10.1111/j.1365-2958.1995.mmi_17030555.x. [DOI] [PubMed] [Google Scholar]
  40. Weiser J. N., Markiewicz Z., Tuomanen E. I., Wani J. H. Relationship between phase variation in colony morphology, intrastrain variation in cell wall physiology, and nasopharyngeal colonization by Streptococcus pneumoniae. Infect Immun. 1996 Jun;64(6):2240–2245. doi: 10.1128/iai.64.6.2240-2245.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. von Heijne G. A new method for predicting signal sequence cleavage sites. Nucleic Acids Res. 1986 Jun 11;14(11):4683–4690. doi: 10.1093/nar/14.11.4683. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES