Abstract
Although more than 98% of natural isolates of Haemophilus influenzae type b carry a duplication of 17 kilobases (kb) of DNA at the chromosomal capsulation locus, only one copy is required for capsulation. In one laboratory-derived and two clinical type b strains, the capsulation locus had a single copy of this 17-kb segment, together with 1.3 kb of DNA identified as lying between the repeats of the duplicated locus. This 1.3 kb appears to be crucial for capsule production, since strains lacking it, although retaining a 17-kb segment, were capsule deficient. On comparing capsule polysaccharide production by these three type b strains with that by a prototypic type b strain with a duplicated locus, a gene dosage effect was demonstrated, with a halving of detectable polysaccharide in the single-copy strains. Despite this reduction in polysaccharide, these strains retained virulence potential as evidenced by bacteremia and meningitis in infant rats. As well as subserving augmented capsule polysaccharide production, a duplicated configuration of the type b cap locus endows strains with genetic instability not found in capsulate single-copy variants. We speculate that a survival advantage might be conferred on strains carrying a duplication at this locus as a result of gene dosage, the genetic instability of the locus, or both.
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- Allan I., Loeb M. R., Moxon E. R. Limited genetic diversity of Haemophilus influenzae (type b). Microb Pathog. 1987 Feb;2(2):139–145. doi: 10.1016/0882-4010(87)90105-7. [DOI] [PubMed] [Google Scholar]
- Anderson P., Johnston R. B., Jr, Smith D. H. Human serum activities against Hemophilus influenzae, type b. J Clin Invest. 1972 Jan;51(1):31–38. doi: 10.1172/JCI106793. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Anderson P., Smith D. H. Isolation of the capsular polysaccharide from culture supernatant of Haemophilus influenzae type b. Infect Immun. 1977 Feb;15(2):472–477. doi: 10.1128/iai.15.2.472-477.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Boulnois G. J., Roberts I. S., Hodge R., Hardy K. R., Jann K. B., Timmis K. N. Analysis of the K1 capsule biosynthesis genes of Escherichia coli: definition of three functional regions for capsule production. Mol Gen Genet. 1987 Jun;208(1-2):242–246. doi: 10.1007/BF00330449. [DOI] [PubMed] [Google Scholar]
- Catlin B. W. Haemophilus influenzae in cultures of cerebrospinal fluid. Noncapsulated variants typable by immunofluorescence. Am J Dis Child. 1970 Sep;120(3):203–210. doi: 10.1001/archpedi.1970.02100080087005. [DOI] [PubMed] [Google Scholar]
- Crisel R. M., Baker R. S., Dorman D. E. Capsular polymer of Haemophilus influenzae, type b. I. Structural characterization of the capsular polymer of strain Eagan. J Biol Chem. 1975 Jul 10;250(13):4926–4930. [PubMed] [Google Scholar]
- Ely S., Tippett J., Kroll J. S., Moxon E. R. Mutations affecting expression and maintenance of genes encoding the serotype b capsule of Haemophilus influenzae. J Bacteriol. 1986 Jul;167(1):44–48. doi: 10.1128/jb.167.1.44-48.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Herriott R. M., Meyer E. M., Vogt M. Defined nongrowth media for stage II development of competence in Haemophilus influenzae. J Bacteriol. 1970 Feb;101(2):517–524. doi: 10.1128/jb.101.2.517-524.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hoiseth S. K., Connelly C. J., Moxon E. R. Genetics of spontaneous, high-frequency loss of b capsule expression in Haemophilus influenzae. Infect Immun. 1985 Aug;49(2):389–395. doi: 10.1128/iai.49.2.389-395.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hoiseth S. K., Moxon E. R., Silver R. P. Genes involved in Haemophilus influenzae type b capsule expression are part of an 18-kilobase tandem duplication. Proc Natl Acad Sci U S A. 1986 Feb;83(4):1106–1110. doi: 10.1073/pnas.83.4.1106. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 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]
- Moxon E. R., Vaughn K. A. The type b capsular polysaccharide as a virulence determinant of Haemophilus influenzae: studies using clinical isolates and laboratory transformants. J Infect Dis. 1981 Apr;143(4):517–524. doi: 10.1093/infdis/143.4.517. [DOI] [PubMed] [Google Scholar]
- Pepple J., Moxon E. R., Yolken R. H. Indirect enzyme-linked immunosorbent assay for the quantitation of the type-specific antigen of Haemophilus influenzae b: a preliminary report. J Pediatr. 1980 Aug;97(2):233–237. doi: 10.1016/s0022-3476(80)80480-x. [DOI] [PubMed] [Google Scholar]
- 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]
- Rigby P. W., Dieckmann M., Rhodes C., Berg P. Labeling deoxyribonucleic acid to high specific activity in vitro by nick translation with DNA polymerase I. J Mol Biol. 1977 Jun 15;113(1):237–251. doi: 10.1016/0022-2836(77)90052-3. [DOI] [PubMed] [Google Scholar]
- Rubin L. G., Moxon E. R. Pathogenesis of bloodstream invasion with Haemophilus influenzae type b. Infect Immun. 1983 Jul;41(1):280–284. doi: 10.1128/iai.41.1.280-284.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Smith A. L., Smith D. H., Averill D. R., Jr, Marino J., Moxon E. R. Production of Haemophilus influenzae b meningitis in infant rats by intraperitoneal inoculation. Infect Immun. 1973 Aug;8(2):278–290. doi: 10.1128/iai.8.2.278-290.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Smith G. E., Summers M. D. The bidirectional transfer of DNA and RNA to nitrocellulose or diazobenzyloxymethyl-paper. Anal Biochem. 1980 Nov 15;109(1):123–129. doi: 10.1016/0003-2697(80)90019-6. [DOI] [PubMed] [Google Scholar]
- Sutton A., Schneerson R., Kendall-Morris S., Robbins J. B. Differential complement resistance mediates virulence of Haemophilus influenzae type b. Infect Immun. 1982 Jan;35(1):95–104. doi: 10.1128/iai.35.1.95-104.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Zwahlen A., Rubin L. G., Moxon E. R. Contribution of lipopolysaccharide to pathogenicity of Haemophilus influenzae: comparative virulence of genetically-related strains in rats. Microb Pathog. 1986 Oct;1(5):465–473. doi: 10.1016/0882-4010(86)90008-2. [DOI] [PubMed] [Google Scholar]