Search tips
Search criteria 


Logo of jbacterPermissionsJournals.ASM.orgJournalJB ArticleJournal InfoAuthorsReviewers
J Bacteriol. 1989 February; 171(2): 722–730.
PMCID: PMC209657

Molecular characterization and protein analysis of the cap region, which is essential for encapsulation in Bacillus anthracis.


By using genetic complementation tests with various in vitro-constructed mutants with mutations in the cap region (which is essential for encapsulation in Bacillus anthracis), we identified three cistrons, capB, capC, and capA, in this order of arrangement. Minicell analysis revealed that these cistrons produce proteins of 44, 16, and 46 kilodaltons, respectively. The complete nucleotide sequence of 3,244 base pairs covering the whole cap region was determined and revealed the existence of the three open reading frames of capB (397 amino acid residues; molecular weight, 44,872), capC (149 amino acid residues; molecular weight, 16,522), and capA (411 amino acid residues; molecular weight, 46,420) arranged in the order predicted by complementation tests. These three cistrons were all transcribed in the same direction from promoters unique to each cistron. Judging from the predicted amino acid sequence of the three proteins and from their localization and their sensitivity to various physicochemical treatments, they appeared to be membrane-associated enzymes mediating the polymerization of D-glutamic acid via the membrane. Capsular peptides immunologically identical to that of B. anthracis were found in B. subtilis, B. megaterium, and B. licheniformis, but no sequence homologous to the cap region was found in any of these bacilli other than B. anthracis. Using strains of B. anthracis with or without insertional inactivation of the cap region, we found that the capsule of B. anthracis conferred strong resistance to phagocytosis upon the bacterial host.

Full text

Full text is available as a scanned copy of the original print version. Get a printable copy (PDF file) of the complete article (2.0M), or click on a page image below to browse page by page. Links to PubMed are also available for Selected References.

Images in this article

Click on the image to see a larger version.

Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • Beachey EH, Ofek I. Epithelial cell binding of group A streptococci by lipoteichoic acid on fimbriae denuded of M protein. J Exp Med. 1976 Apr 1;143(4):759–771. [PMC free article] [PubMed]
  • BEALL FA, TAYLOR MJ, THORNE CB. Rapid lethal effect in rats of a third component found upon fractionating the toxin of Bacillus anthracis. J Bacteriol. 1962 Jun;83:1274–1280. [PMC free article] [PubMed]
  • Bolivar F, Rodriguez RL, Greene PJ, Betlach MC, Heyneker HL, Boyer HW, Crosa JH, Falkow S. Construction and characterization of new cloning vehicles. II. A multipurpose cloning system. Gene. 1977;2(2):95–113. [PubMed]
  • Brosius J. Plasmid vectors for the selection of promoters. Gene. 1984 Feb;27(2):151–160. [PubMed]
  • Casadaban MJ, Cohen SN. Analysis of gene control signals by DNA fusion and cloning in Escherichia coli. J Mol Biol. 1980 Apr;138(2):179–207. [PubMed]
  • Gardner JM, Troy FA. Chemistry and biosynthesis of the poly(gamma-D-glutamyl) capsule in Bacillus licheniformis. Activation, racemization, and polymerization of glutamic acid by a membranous polyglutamyl synthetase complex. J Biol Chem. 1979 Jul 25;254(14):6262–6269. [PubMed]
  • Green BD, Battisti L, Koehler TM, Thorne CB, Ivins BE. Demonstration of a capsule plasmid in Bacillus anthracis. Infect Immun. 1985 Aug;49(2):291–297. [PMC free article] [PubMed]
  • GUEX-HOLZER S, TOMCSIK J. The isolation and chemical nature of capsular and cell-wall haptens in a Bacillus species. J Gen Microbiol. 1956 Feb;14(1):14–25. [PubMed]
  • Hanby WE, Rydon HN. The capsular substance of Bacillus anthracis: With an appendix by P. Bruce White. Biochem J. 1946;40(2):297–309. [PubMed]
  • Ishiwa H, Tsuchida N. New shuttle vectors for Escherichia coli and Bacillus subtilis. I. Construction and characterization of plasmid pHY460 with twelve unique cloning sites. Gene. 1984 Dec;32(1-2):129–134. [PubMed]
  • Ito K, Sato T, Yura T. Synthesis and assembly of the membrane proteins in E. coli. Cell. 1977 Jul;11(3):551–559. [PubMed]
  • Jones BD, Mobley HL. Genetic and biochemical diversity of ureases of Proteus, Providencia, and Morganella species isolated from urinary tract infection. Infect Immun. 1987 Sep;55(9):2198–2203. [PMC free article] [PubMed]
  • Klein P, Kanehisa M, DeLisi C. The detection and classification of membrane-spanning proteins. Biochim Biophys Acta. 1985 May 28;815(3):468–476. [PubMed]
  • Kyte J, Doolittle RF. A simple method for displaying the hydropathic character of a protein. J Mol Biol. 1982 May 5;157(1):105–132. [PubMed]
  • Liu TY, Gotschlich EC, Jonssen EK, Wysocki JR. Studies on the meningococcal polysaccharides. I. Composition and chemical properties of the group A polysaccharide. J Biol Chem. 1971 May 10;246(9):2849–2858. [PubMed]
  • Makino S, Sasakawa C, Kamata K, Kurata T, Yoshikawa M. A genetic determinant required for continuous reinfection of adjacent cells on large plasmid in S. flexneri 2a. Cell. 1986 Aug 15;46(4):551–555. [PubMed]
  • Makino S, Sasakawa C, Uchida I, Terakado N, Yoshikawa M. Cloning and CO2-dependent expression of the genetic region for encapsulation from Bacillus anthracis. Mol Microbiol. 1988 May;2(3):371–376. [PubMed]
  • Manoil C, Beckwith J. TnphoA: a transposon probe for protein export signals. Proc Natl Acad Sci U S A. 1985 Dec;82(23):8129–8133. [PubMed]
  • Mikesell P, Ivins BE, Ristroph JD, Dreier TM. Evidence for plasmid-mediated toxin production in Bacillus anthracis. Infect Immun. 1983 Jan;39(1):371–376. [PMC free article] [PubMed]
  • Nakayama R, Kumagai H, Tochikura T. Leakage of glutathione from bacterial cells caused by inhibition of gamma-glutamyltranspeptidase. Appl Environ Microbiol. 1984 Apr;47(4):653–657. [PMC free article] [PubMed]
  • Osborn MJ. Structure and biosynthesis of the bacterial cell wall. Annu Rev Biochem. 1969;38:501–538. [PubMed]
  • Sanger F, Nicklen S, Coulson AR. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. [PubMed]
  • Sasakawa C, Yoshikawa M. A series of Tn5 variants with various drug-resistance markers and suicide vector for transposon mutagenesis. Gene. 1987;56(2-3):283–288. [PubMed]
  • Scher M, Lennarz WJ. Studies on the biosynthesis of mannan in Micrococcus lysodeikticus. I. Characterization of mannan-14C formed enzymatically from mannosyl-1-phosphoryl-undecaprenol. J Biol Chem. 1969 May 25;244(10):2777–2789. [PubMed]
  • Shiba K, Ito K, Yura T, Cerretti DP. A defined mutation in the protein export gene within the spc ribosomal protein operon of Escherichia coli: isolation and characterization of a new temperature-sensitive secY mutant. EMBO J. 1984 Mar;3(3):631–635. [PubMed]
  • 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. [PubMed]
  • STANLEY JL, SARGEANT K, SMITH H. Purification of factors I and II of the anthrax toxin produced in vivo. J Gen Microbiol. 1960 Feb;22:206–218. [PubMed]
  • Suzuki H, Kumagai H, Tochikura T. Isolation, genetic mapping, and characterization of Escherichia coli K-12 mutants lacking gamma-glutamyltranspeptidase. J Bacteriol. 1987 Sep;169(9):3926–3931. [PMC free article] [PubMed]
  • Thompson R, Achtman M. The control region of the F sex factor DNA transfer cistrons: restriction mapping and DNA cloning. Mol Gen Genet. 1978 Oct 24;165(3):295–304. [PubMed]
  • THORNE CB, GOMEZ CG, HOUSEWRIGHT RD. Transamination of D-amino acids by Bacillus subtilis. J Bacteriol. 1955 Mar;69(3):357–362. [PMC free article] [PubMed]
  • THORNE CB, LEONARD CG. Isolation of D- and L-glutamyl polypeptides from culture filtrates of Bacillus subtilis. J Biol Chem. 1958 Nov;233(5):1109–1112. [PubMed]
  • THORNE CB, MOLNAR DM. D-Amino acid transamination in bacillus anthracis. J Bacteriol. 1955 Oct;70(4):420–426. [PMC free article] [PubMed]
  • THORNE CB, MOLNAR DM, STRANGE RE. Production of toxin in vitro by Bacillus anthracis and its spearation into two components. J Bacteriol. 1960 Mar;79:450–455. [PMC free article] [PubMed]
  • Troy FA. Chemistry and biosynthesis of the poly( -D-glutamyl) capsule in Bacillus licheniformis. I. Properties of the membrane-mediated biosynthetic reaction. J Biol Chem. 1973 Jan 10;248(1):305–315. [PubMed]
  • Uchida I, Hashimoto K, Makino S, Sasakawa C, Yoshikawa M, Terakado N. Restriction map of a capsule plasmid of Bacillus anthracis. Plasmid. 1987 Sep;18(2):178–181. [PubMed]
  • Uchida I, Sekizaki T, Hashimoto K, Terakado N. Association of the encapsulation of Bacillus anthracis with a 60 megadalton plasmid. J Gen Microbiol. 1985 Feb;131(2):363–367. [PubMed]
  • Vijay IK, Troy FA. Properties of membrane-associated sialyltransferase of Escherichia coli. J Biol Chem. 1975 Jan 10;250(1):164–170. [PubMed]

Articles from Journal of Bacteriology are provided here courtesy of American Society for Microbiology (ASM)