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Infect Immun. Dec 1994; 62(12): 5384–5396.
PMCID: PMC303279

Nucleotide sequence analysis of genes essential for capsular polysaccharide biosynthesis in Streptococcus pneumoniae type 19F.

Abstract

Previous studies have shown that the capsular polysaccharide synthesis (cps) locus of the type 19F Streptococcus pneumoniae strain SSZ was closely linked to a copy of the insertion sequence IS1202 (J.K. Morona, A. Guidolin, R. Morona, D. Hansman, and J.C. Paton, J. Bacteriol. 176:4437-4443, 1994). In the present study, we used plasmid insertion and rescue and inverse PCR to clone 6,322 bp of flanking DNA upstream of IS1202. Sequence analysis indicated that this region contains six complete open reading frames (ORFs) and one partial ORF that are arranged as a single transcriptional unit. Chromosomal disruption of any of these ORFs in a smooth-type 19F strain leads to a rough (unencapsulated) phenotype, indicating that this operon is essential for capsule production. The ORFs have therefore been designated cps19fA to cps19fG, where cps19fA is the first gene of the type 19F cps locus. Furthermore, many of the gene products from this incomplete operon exhibit strong similarities to proteins known to be involved in the production of capsular polysaccharide, exopolysaccharide, teichoic acid, enterobacterial common antigen, and lipopolysaccharide from numerous other bacterial species. This has allowed us to propose functions for many of the type 19F cps gene products. Southern hybridization studies reveal that cps19fA and cps19fB are conserved among all 12 pneumococcal serotypes tested, whereas genes downstream of cps19fB are conserved among some, but not all, of the serotypes tested.

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  • Altschul SF, Gish W, Miller W, Myers EW, Lipman DJ. Basic local alignment search tool. J Mol Biol. 1990 Oct 5;215(3):403–410. [PubMed]
  • Austrian R. Some observations on the pneumococcus and on the current status of pneumococcal disease and its prevention. Rev Infect Dis. 1981 Mar-Apr;3 (Suppl):S1–17. [PubMed]
  • AUSTRIAN R, BERNHEIMER HP, SMITH EE, MILLS GT. Simultaneous production of two capsular polysaccharides by pneumococcus. II. The genetic and biochemical bases of binary capsulation. J Exp Med. 1959 Oct 1;110:585–602. [PMC free article] [PubMed]
  • Berry AM, Yother J, Briles DE, Hansman D, Paton JC. Reduced virulence of a defined pneumolysin-negative mutant of Streptococcus pneumoniae. Infect Immun. 1989 Jul;57(7):2037–2042. [PMC free article] [PubMed]
  • Borthakur D, Barker RF, Latchford JW, Rossen L, Johnston AW. Analysis of pss genes of Rhizobium leguminosarum required for exopolysaccharide synthesis and nodulation of peas: their primary structure and their interaction with psi and other nodulation genes. Mol Gen Genet. 1988 Jul;213(1):155–162. [PubMed]
  • Daniels DL, Plunkett G, 3rd, Burland V, Blattner FR. Analysis of the Escherichia coli genome: DNA sequence of the region from 84.5 to 86.5 minutes. Science. 1992 Aug 7;257(5071):771–778. [PubMed]
  • Dillard JP, Yother J. Genetic and molecular characterization of capsular polysaccharide biosynthesis in Streptococcus pneumoniae type 3. Mol Microbiol. 1994 Jun;12(6):959–972. [PubMed]
  • Douglas RM, Paton JC, Duncan SJ, Hansman DJ. Antibody response to pneumococcal vaccination in children younger than five years of age. J Infect Dis. 1983 Jul;148(1):131–137. [PubMed]
  • Frosch M, Weisgerber C, Meyer TF. Molecular characterization and expression in Escherichia coli of the gene complex encoding the polysaccharide capsule of Neisseria meningitidis group B. Proc Natl Acad Sci U S A. 1989 Mar;86(5):1669–1673. [PMC free article] [PubMed]
  • García E, García P, López R. Cloning and sequencing of a gene involved in the synthesis of the capsular polysaccharide of Streptococcus pneumoniae type 3. Mol Gen Genet. 1993 May;239(1-2):188–195. [PubMed]
  • Glucksmann MA, Reuber TL, Walker GC. Family of glycosyl transferases needed for the synthesis of succinoglycan by Rhizobium meliloti. J Bacteriol. 1993 Nov;175(21):7033–7044. [PMC free article] [PubMed]
  • Glucksmann MA, Reuber TL, Walker GC. Genes needed for the modification, polymerization, export, and processing of succinoglycan by Rhizobium meliloti: a model for succinoglycan biosynthesis. J Bacteriol. 1993 Nov;175(21):7045–7055. [PMC free article] [PubMed]
  • Gray BM, Converse GM, 3rd, Dillon HC., Jr Epidemiologic studies of Streptococcus pneumoniae in infants: acquisition, carriage, and infection during the first 24 months of life. J Infect Dis. 1980 Dec;142(6):923–933. [PubMed]
  • Gray JX, Djordjevic MA, Rolfe BG. Two genes that regulate exopolysaccharide production in Rhizobium sp. strain NGR234: DNA sequences and resultant phenotypes. J Bacteriol. 1990 Jan;172(1):193–203. [PMC free article] [PubMed]
  • Hanahan D. Studies on transformation of Escherichia coli with plasmids. J Mol Biol. 1983 Jun 5;166(4):557–580. [PubMed]
  • Henikoff S. Unidirectional digestion with exonuclease III creates targeted breakpoints for DNA sequencing. Gene. 1984 Jun;28(3):351–359. [PubMed]
  • Higgins DG, Sharp PM. CLUSTAL: a package for performing multiple sequence alignment on a microcomputer. Gene. 1988 Dec 15;73(1):237–244. [PubMed]
  • Ielpi L, Couso RO, Dankert MA. Sequential assembly and polymerization of the polyprenol-linked pentasaccharide repeating unit of the xanthan polysaccharide in Xanthomonas campestris. J Bacteriol. 1993 May;175(9):2490–2500. [PMC free article] [PubMed]
  • Jiang XM, Neal B, Santiago F, Lee SJ, Romana LK, Reeves PR. Structure and sequence of the rfb (O antigen) gene cluster of Salmonella serovar typhimurium (strain LT2). Mol Microbiol. 1991 Mar;5(3):695–713. [PubMed]
  • Kroll JS, Zamze S, Loynds B, Moxon ER. Common organization of chromosomal loci for production of different capsular polysaccharides in Haemophilus influenzae. J Bacteriol. 1989 Jun;171(6):3343–3347. [PMC free article] [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]
  • Lazarevic V, Margot P, Soldo B, Karamata D. Sequencing and analysis of the Bacillus subtilis lytRABC divergon: a regulatory unit encompassing the structural genes of the N-acetylmuramoyl-L-alanine amidase and its modifier. J Gen Microbiol. 1992 Sep;138(9):1949–1961. [PubMed]
  • Lee CJ, Banks SD, Li JP. Virulence, immunity, and vaccine related to Streptococcus pneumoniae. Crit Rev Microbiol. 1991;18(2):89–114. [PubMed]
  • Lu CD, Abdelal AT. The Salmonella typhimurium uracil-sensitive mutation use is in argU and encodes a minor arginine tRNA. J Bacteriol. 1993 Jun;175(12):3897–3899. [PMC free article] [PubMed]
  • Macrina FL, Evans RP, Tobian JA, Hartley DL, Clewell DB, Jones KR. Novel shuttle plasmid vehicles for Escherichia-Streptococcus transgeneric cloning. Gene. 1983 Nov;25(1):145–150. [PubMed]
  • Mauël C, Young M, Karamata D. Genes concerned with synthesis of poly(glycerol phosphate), the essential teichoic acid in Bacillus subtilis strain 168, are organized in two divergent transcription units. J Gen Microbiol. 1991 Apr;137(4):929–941. [PubMed]
  • Morona JK, Guidolin A, Morona R, Hansman D, Paton JC. Isolation, characterization, and nucleotide sequence of IS1202, an insertion sequence of Streptococcus pneumoniae. J Bacteriol. 1994 Jul;176(14):4437–4443. [PMC free article] [PubMed]
  • Müller P, Keller M, Weng WM, Quandt J, Arnold W, Pühler A. Genetic analysis of the Rhizobium meliloti exoYFQ operon: ExoY is homologous to sugar transferases and ExoQ represents a transmembrane protein. Mol Plant Microbe Interact. 1993 Jan-Feb;6(1):55–65. [PubMed]
  • Musher DM. Infections caused by Streptococcus pneumoniae: clinical spectrum, pathogenesis, immunity, and treatment. Clin Infect Dis. 1992 Apr;14(4):801–807. [PubMed]
  • Ochman H, Gerber AS, Hartl DL. Genetic applications of an inverse polymerase chain reaction. Genetics. 1988 Nov;120(3):621–623. [PMC free article] [PubMed]
  • Paton JC, Berry AM, Lock RA, Hansman D, Manning PA. Cloning and expression in Escherichia coli of the Streptococcus pneumoniae gene encoding pneumolysin. Infect Immun. 1986 Oct;54(1):50–55. [PMC free article] [PubMed]
  • Reed JW, Capage M, Walker GC. Rhizobium meliloti exoG and exoJ mutations affect the exoX-exoY system for modulation of exopolysaccharide production. J Bacteriol. 1991 Jun;173(12):3776–3788. [PMC free article] [PubMed]
  • Reuber TL, Walker GC. Biosynthesis of succinoglycan, a symbiotically important exopolysaccharide of Rhizobium meliloti. Cell. 1993 Jul 30;74(2):269–280. [PubMed]
  • Roberts IS, Mountford R, Hodge R, Jann KB, Boulnois GJ. Common organization of gene clusters for production of different capsular polysaccharides (K antigens) in Escherichia coli. J Bacteriol. 1988 Mar;170(3):1305–1310. [PMC free article] [PubMed]
  • Rubens CE, Heggen LM, Haft RF, Wessels MR. Identification of cpsD, a gene essential for type III capsule expression in group B streptococci. Mol Microbiol. 1993 May;8(5):843–855. [PubMed]
  • Shoemaker NB, Guild WR. Destruction of low efficiency markers is a slow process occurring at a heteroduplex stage of transformation. Mol Gen Genet. 1974;128(4):283–290. [PubMed]
  • Southern EM. Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol. 1975 Nov 5;98(3):503–517. [PubMed]
  • Wang L, Reeves PR. Involvement of the galactosyl-1-phosphate transferase encoded by the Salmonella enterica rfbP gene in O-antigen subunit processing. J Bacteriol. 1994 Jul;176(14):4348–4356. [PMC free article] [PubMed]
  • Watson DA, Musher DM. Interruption of capsule production in Streptococcus pneumonia serotype 3 by insertion of transposon Tn916. Infect Immun. 1990 Sep;58(9):3135–3138. [PMC free article] [PubMed]
  • Weiser JN, Love JM, Moxon ER. The molecular mechanism of phase variation of H. influenzae lipopolysaccharide. Cell. 1989 Nov 17;59(4):657–665. [PubMed]
  • Yother J, McDaniel LS, Briles DE. Transformation of encapsulated Streptococcus pneumoniae. J Bacteriol. 1986 Dec;168(3):1463–1465. [PMC free article] [PubMed]

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