Skip to main content
PMC home page
Infection and Immunity logoLink to Infection and Immunity
. 1996 Oct;64(10):4000–4007. doi: 10.1128/iai.64.10.4000-4007.1996

The hemagglutinin gene A (hagA) of Porphyromonas gingivalis 381 contains four large, contiguous, direct repeats.

N Han 1, J Whitlock 1, A Progulske-Fox 1
PMCID: PMC174329  PMID: 8926061

Abstract

Porphyromonas gingivalis is a gram-negative anaerobic bacterial species strongly associated with adult periodontitis. One of its distinguishing characteristics and putative virulence properties is the ability to agglutinate erythrocytes. We have previously reported the cloning of multiple hemagglutinin genes from P. gingivalis 381. Subsequent sequencing of clone ST 2 revealed that the cloned fragment contained only an internal portion of the gene which lacked both start and stop codons. We here report the cloning and sequencing of the entire gene, designated hagA, as well as its relationship to other genes of this species. By use of inverse PCR technology and the construction of several additional genomic libraries, the complete open reading frame of hagA was found to be 7,887 bp in length, encoding a protein of 2,628 amino acids with a molecular mass of 283.3 kDa, which is among the largest genes ever cloned from a prokaryote to date. Within its open reading frame, four large, contiguous, direct repeats (varying from 1,318 to 1,368 bp) were identified. The repeat unit (HArep), which is assumed to contain the hemagglutinin domain, is also present in other recently reported protease and hemagglutinin genes in P. gingivalis. Thus, we propose that hagA and the other genes which share the HArep sequence form a multigene family with hagA as a central member.

Full Text

The Full Text of this article is available as a PDF (440.1 KB).

Selected References

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

  1. Allred D. R., McGuire T. C., Palmer G. H., Leib S. R., Harkins T. M., McElwain T. F., Barbet A. F. Molecular basis for surface antigen size polymorphisms and conservation of a neutralization-sensitive epitope in Anaplasma marginale. Proc Natl Acad Sci U S A. 1990 Apr;87(8):3220–3224. doi: 10.1073/pnas.87.8.3220. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Anderson B. E., McDonald G. A., Jones D. C., Regnery R. L. A protective protein antigen of Rickettsia rickettsii has tandemly repeated, near-identical sequences. Infect Immun. 1990 Sep;58(9):2760–2769. doi: 10.1128/iai.58.9.2760-2769.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Aslanzadeh J., Paulissen L. J. Adherence and pathogenesis of Salmonella enteritidis in mice. Microbiol Immunol. 1990;34(11):885–893. doi: 10.1111/j.1348-0421.1990.tb01067.x. [DOI] [PubMed] [Google Scholar]
  4. Booth B. A., Boesman-Finkelstein M., Finkelstein R. A. Vibrio cholerae hemagglutinin/protease nicks cholera enterotoxin. Infect Immun. 1984 Sep;45(3):558–560. doi: 10.1128/iai.45.3.558-560.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Bourgeau G., Lapointe H., Péloquin P., Mayrand D. Cloning, expression, and sequencing of a protease gene (tpr) from Porphyromonas gingivalis W83 in Escherichia coli. Infect Immun. 1992 Aug;60(8):3186–3192. doi: 10.1128/iai.60.8.3186-3192.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Brennan M. J., Li Z. M., Cowell J. L., Bisher M. E., Steven A. C., Novotny P., Manclark C. R. Identification of a 69-kilodalton nonfimbrial protein as an agglutinogen of Bordetella pertussis. Infect Immun. 1988 Dec;56(12):3189–3195. doi: 10.1128/iai.56.12.3189-3195.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Caspers P., Gentz R., Matile H., Pink J. R., Sinigaglia F. The circumsporozoite protein gene from NF54, a Plasmodium falciparum isolate used in malaria vaccine trials. Mol Biochem Parasitol. 1989 Jun 15;35(2):185–189. doi: 10.1016/0166-6851(89)90121-7. [DOI] [PubMed] [Google Scholar]
  8. Duguid J. P., Anderson E. S., Campbell I. Fimbriae and adhesive properties in Salmonellae. J Pathol Bacteriol. 1966 Jul;92(1):107–138. doi: 10.1002/path.1700920113. [DOI] [PubMed] [Google Scholar]
  9. Duguid J. P., Clegg S., Wilson M. I. The fimbrial and non-fimbrial haemagglutinins of Escherichia coli. J Med Microbiol. 1979 May;12(2):213–227. doi: 10.1099/00222615-12-2-213. [DOI] [PubMed] [Google Scholar]
  10. Fischetti V. A. Streptococcal M protein. Sci Am. 1991 Jun;264(6):58–65. doi: 10.1038/scientificamerican0691-58. [DOI] [PubMed] [Google Scholar]
  11. Fletcher H. M., Schenkein H. A., Macrina F. L. Cloning and characterization of a new protease gene (prtH) from Porphyromonas gingivalis. Infect Immun. 1994 Oct;62(10):4279–4286. doi: 10.1128/iai.62.10.4279-4286.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Hoover C. I., Ng C. Y., Felton J. R. Correlation of haemagglutination activity with trypsin-like protease activity of Porphyromonas gingivalis. Arch Oral Biol. 1992;37(7):515–520. doi: 10.1016/0003-9969(92)90133-s. [DOI] [PubMed] [Google Scholar]
  13. Jameson B. A., Wolf H. The antigenic index: a novel algorithm for predicting antigenic determinants. Comput Appl Biosci. 1988 Mar;4(1):181–186. doi: 10.1093/bioinformatics/4.1.181. [DOI] [PubMed] [Google Scholar]
  14. Kato T., Takahashi N., Kuramitsu H. K. Sequence analysis and characterization of the Porphyromonas gingivalis prtC gene, which expresses a novel collagenase activity. J Bacteriol. 1992 Jun;174(12):3889–3895. doi: 10.1128/jb.174.12.3889-3895.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Kirszbaum L., Sotiropoulos C., Jackson C., Cleal S., Slakeski N., Reynolds E. C. Complete nucleotide sequence of a gene prtR of Porphyromonas gingivalis W50 encoding a 132 kDa protein that contains an arginine-specific thiol endopeptidase domain and a haemagglutinin domain. Biochem Biophys Res Commun. 1995 Feb 6;207(1):424–431. doi: 10.1006/bbrc.1995.1205. [DOI] [PubMed] [Google Scholar]
  16. Ladefoged S. A., Birkelund S., Hauge S., Brock B., Jensen L. T., Christiansen G. A 135-kilodalton surface antigen of Mycoplasma hominis PG21 contains multiple directly repeated sequences. Infect Immun. 1995 Jan;63(1):212–223. doi: 10.1128/iai.63.1.212-223.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Lépine G., Progulske-Fox A. Duplication and differential expression of hemagglutinin genes in Porphyromonas gingivalis. Oral Microbiol Immunol. 1996 Apr;11(2):65–78. doi: 10.1111/j.1399-302x.1996.tb00339.x. [DOI] [PubMed] [Google Scholar]
  18. Madden T. E., Clark V. L., Kuramitsu H. K. Revised sequence of the Porphyromonas gingivalis prtT cysteine protease/hemagglutinin gene: homology with streptococcal pyrogenic exotoxin B/streptococcal proteinase. Infect Immun. 1995 Jan;63(1):238–247. doi: 10.1128/iai.63.1.238-247.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Markham P. F., Glew M. D., Whithear K. G., Walker I. D. Molecular cloning of a member of the gene family that encodes pMGA, a hemagglutinin of Mycoplasma gallisepticum. Infect Immun. 1993 Mar;61(3):903–909. doi: 10.1128/iai.61.3.903-909.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. McNab R., Jenkinson H. F., Loach D. M., Tannock G. W. Cell-surface-associated polypeptides CshA and CshB of high molecular mass are colonization determinants in the oral bacterium Streptococcus gordonii. Mol Microbiol. 1994 Nov;14(4):743–754. doi: 10.1111/j.1365-2958.1994.tb01311.x. [DOI] [PubMed] [Google Scholar]
  21. Nishikata M., Yoshimura F. Characterization of Porphyromonas (bacteroides) gingivalis hemagglutinin as a protease. Biochem Biophys Res Commun. 1991 Jul 15;178(1):336–342. doi: 10.1016/0006-291x(91)91819-x. [DOI] [PubMed] [Google Scholar]
  22. Okamoto K., Misumi Y., Kadowaki T., Yoneda M., Yamamoto K., Ikehara Y. Structural characterization of argingipain, a novel arginine-specific cysteine proteinase as a major periodontal pathogenic factor from Porphyromonas gingivalis. Arch Biochem Biophys. 1995 Feb 1;316(2):917–925. doi: 10.1006/abbi.1995.1123. [DOI] [PubMed] [Google Scholar]
  23. Okuda K., Takazoe I. Haemagglutinating activity of Bacteroides melaninogenicus. Arch Oral Biol. 1974 May;19(5):415–416. doi: 10.1016/0003-9969(74)90184-8. [DOI] [PubMed] [Google Scholar]
  24. Pavloff N., Potempa J., Pike R. N., Prochazka V., Kiefer M. C., Travis J., Barr P. J. Molecular cloning and structural characterization of the Arg-gingipain proteinase of Porphyromonas gingivalis. Biosynthesis as a proteinase-adhesin polyprotein. J Biol Chem. 1995 Jan 20;270(3):1007–1010. doi: 10.1074/jbc.270.3.1007. [DOI] [PubMed] [Google Scholar]
  25. Pike R., McGraw W., Potempa J., Travis J. Lysine- and arginine-specific proteinases from Porphyromonas gingivalis. Isolation, characterization, and evidence for the existence of complexes with hemagglutinins. J Biol Chem. 1994 Jan 7;269(1):406–411. [PubMed] [Google Scholar]
  26. Progulske-Fox A., Rao V., Han N., Lepine G., Witlock J., Lantz M. Molecular characterization of hemagglutinin genes of periodontopathic bacteria. J Periodontal Res. 1993 Nov;28(6 Pt 2):473–474. doi: 10.1111/j.1600-0765.1993.tb02106.x. [DOI] [PubMed] [Google Scholar]
  27. Progulske-Fox A., Tumwasorn S., Holt S. C. The expression and function of a Bacteroides gingivalis hemagglutinin gene in Escherichia coli. Oral Microbiol Immunol. 1989 Sep;4(3):121–131. doi: 10.1111/j.1399-302x.1989.tb00238.x. [DOI] [PubMed] [Google Scholar]
  28. Progulske-Fox A., Tumwasorn S., Lépine G., Whitlock J., Savett D., Ferretti J. J., Banas J. A. The cloning, expression and sequence analysis of a second Porphyromonas gingivalis gene that codes for a protein involved in hemagglutination. Oral Microbiol Immunol. 1995 Oct;10(5):311–318. doi: 10.1111/j.1399-302x.1995.tb00160.x. [DOI] [PubMed] [Google Scholar]
  29. 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]
  30. Reuven N. B., Koonin E. V., Rudd K. E., Deutscher M. P. The gene for the longest known Escherichia coli protein is a member of helicase superfamily II. J Bacteriol. 1995 Oct;177(19):5393–5400. doi: 10.1128/jb.177.19.5393-5400.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Yogev D., Rosengarten R., Watson-McKown R., Wise K. S. Molecular basis of Mycoplasma surface antigenic variation: a novel set of divergent genes undergo spontaneous mutation of periodic coding regions and 5' regulatory sequences. EMBO J. 1991 Dec;10(13):4069–4079. doi: 10.1002/j.1460-2075.1991.tb04983.x. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES