• We are sorry, but NCBI web applications do not support your browser and may not function properly. More information
Logo of jbacterPermissionsJournals.ASM.orgJournalJB ArticleJournal InfoAuthorsReviewers
J Bacteriol. Feb 1997; 179(4): 1126–1134.
PMCID: PMC178808

Biological function of the dTDP-rhamnose synthesis pathway in Streptococcus mutans.

Abstract

We have cloned a new gene locus that comprises three genes concerned with the biosynthesis of the serotype c-specific polysaccharide antigen in Streptococcus mutans. The genes encode proteins exhibiting significant homology to the rfbA, rfbB, and rfbD gene products that are involved in the anabolism of dTDP-L-rhamnose from D-glucose-1-phosphate. This anabolism pathway pertains to biosynthesis of the O antigen of lipopolysaccharide in gram-negative bacteria. The cell extract of Escherichia coli expressing each of the cloned genes of S. mutans exhibited enzymatic activity corresponding to the homologous counterpart of the rfb gene products. Rhamnose was not detected in the cell wall preparation purified from the mutant in which each of the three cloned genes was insertionally inactivated. Rabbit antiserum against S. mutans serotype c-specific antigen did not react with the autoclaved extracts from these mutants. These results indicate that the gene products identified in the present study are involved in the dTDP-L-rhamnose synthesis pathway and that the pathway relates to the biosynthesis of the serotype-specific polysaccharide antigen of S. mutans. Southern hybridization analysis revealed that genes homologous to the cloned genes involved in the dTDP-L-rhamnose synthesis pathway were widely distributed in a variety of streptococci. This is the first report of the biological function of the dTDP-rhamnose pathway in streptococci.

Full Text

The Full Text of this article is available as a PDF (1.5M).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • Boyer HW, Roulland-Dussoix D. A complementation analysis of the restriction and modification of DNA in Escherichia coli. J Mol Biol. 1969 May 14;41(3):459–472. [PubMed]
  • Bratthall D. Demonstration of five serological groups of streptococcal strains resembling Streptococcus mutans. Odontol Revy. 1970;21(2):143–152. [PubMed]
  • Coligan JE, Kindt TJ, Krause RM. Structure of the streptococcal groups A, A-variant and C carbohydrates. Immunochemistry. 1978 Nov;15(10-11):755–760. [PubMed]
  • CURTIS SN, KRAUSE RM. ANTIGENIC RELATIONSHIPS BETWEEN GROUPS B AND G STREPTOCOCCI. J Exp Med. 1964 Oct 1;120:629–637. [PMC free article] [PubMed]
  • CURTIS SN, KRAUSE RM. IMMUNOCHEMICAL STUDIES ON THE SPECIFIC CARBOHYDRATE OF GROUP G STREPTOCOCCI. J Exp Med. 1964 Jan 1;119:997–1003. [PMC free article] [PubMed]
  • Dower WJ, Miller JF, Ragsdale CW. High efficiency transformation of E. coli by high voltage electroporation. Nucleic Acids Res. 1988 Jul 11;16(13):6127–6145. [PMC free article] [PubMed]
  • Duclos B, Cortay JC, Bleicher F, Ron EZ, Richaud C, Saint Girons I, Cozzone AJ. Nucleotide sequence of the metA gene encoding homoserine trans-succinylase in Escherichia coli. Nucleic Acids Res. 1989 Apr 11;17(7):2856–2856. [PMC free article] [PubMed]
  • Hamada S, Slade HD. Purification and immunochemical characterization of type e polysaccharide antigen of Streptococcus mutans. Infect Immun. 1976 Jul;14(1):68–76. [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]
  • Hether NW, Campbell PA, Baker LA, Jackson LL. Chemical composition and biological functions of Listeria monocytogenes cell wall preparations. Infect Immun. 1983 Mar;39(3):1114–1121. [PMC free article] [PubMed]
  • Higgins DG, Bleasby AJ, Fuchs R. CLUSTAL V: improved software for multiple sequence alignment. Comput Appl Biosci. 1992 Apr;8(2):189–191. [PubMed]
  • Imai T, Ohta K, Kigawa H, Kanoh H, Taniguchi T, Tobari J. Preparation of high-molecular-weight DNA: application to mycobacterial cells. Anal Biochem. 1994 Nov 1;222(2):479–482. [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]
  • Koga T, Asakawa H, Okahashi N, Takahashi I. Effect of subculturing on expression of a cell-surface protein antigen by Streptococcus mutans. J Gen Microbiol. 1989 Dec;135(12):3199–3207. [PubMed]
  • Kojic M, Vujcic M, Banina A, Cocconcelli P, Cerning J, Topisirovic L. Analysis of exopolysaccharide production by Lactobacillus casei CG11, isolated from cheese. Appl Environ Microbiol. 1992 Dec;58(12):4086–4088. [PMC free article] [PubMed]
  • Kojima N, Araki Y, Ito E. Structural studies on the acidic polysaccharide of Bacillus cereus AHU 1356 cell walls. Eur J Biochem. 1985 May 2;148(3):479–484. [PubMed]
  • Köplin R, Wang G, Hötte B, Priefer UB, Pühler A. A 3.9-kb DNA region of Xanthomonas campestris pv. campestris that is necessary for lipopolysaccharide production encodes a set of enzymes involved in the synthesis of dTDP-rhamnose. J Bacteriol. 1993 Dec;175(24):7786–7792. [PMC free article] [PubMed]
  • Lindquist L, Kaiser R, Reeves PR, Lindberg AA. Purification, characterization and HPLC assay of Salmonella glucose-1-phosphate thymidylyl-transferase from the cloned rfbA gene. Eur J Biochem. 1993 Feb 1;211(3):763–770. [PubMed]
  • Linzer R, Gill K, Slade HD. Chemical composition of Streptococcus mutans type c antigen: comparison to type a, b, and d antigens. J Dent Res. 1976 Jan;55:A109–A115. [PubMed]
  • Linzer R, Reddy MS, Levine MJ. Structural studies of the rhamnose-glucose polysaccharide antigen from Streptococcus sobrinus B13 and 6715-T2. Infect Immun. 1985 Nov;50(2):583–585. [PMC free article] [PubMed]
  • Lipman DJ, Pearson WR. Rapid and sensitive protein similarity searches. Science. 1985 Mar 22;227(4693):1435–1441. [PubMed]
  • Macpherson DF, Manning PA, Morona R. Characterization of the dTDP-rhamnose biosynthetic genes encoded in the rfb locus of Shigella flexneri. Mol Microbiol. 1994 Jan;11(2):281–292. [PubMed]
  • Marolda CL, Valvano MA. Genetic analysis of the dTDP-rhamnose biosynthesis region of the Escherichia coli VW187 (O7:K1) rfb gene cluster: identification of functional homologs of rfbB and rfbA in the rff cluster and correct location of the rffE gene. J Bacteriol. 1995 Oct;177(19):5539–5546. [PMC free article] [PubMed]
  • Méjean V, Salles C, Bullions LC, Bessman MJ, Claverys JP. Characterization of the mutX gene of Streptococcus pneumoniae as a homologue of Escherichia coli mutT, and tentative definition of a catalytic domain of the dGTP pyrophosphohydrolases. Mol Microbiol. 1994 Jan;11(2):323–330. [PubMed]
  • Melo A, Glaser L. The mechanism of 6-deoxyhexose synthesis. II. Conversion of deoxythymidine diphosphate 4-keto-6-deoxy-D-glucose to deoxythymidine diphosphate L-rhamnose. J Biol Chem. 1968 Apr 10;243(7):1475–1478. [PubMed]
  • Morris DL. Quantitative Determination of Carbohydrates With Dreywood's Anthrone Reagent. Science. 1948 Mar 5;107(2775):254–255. [PubMed]
  • Nagaoka M, Muto M, Nomoto K, Matuzaki T, Watanabe T, Yokokura T. Structure of polysaccharide-peptidoglycan complex from the cell wall of Lactobacillus casei YIT9018. J Biochem. 1990 Oct;108(4):568–571. [PubMed]
  • Niaudet B, Goze A, Ehrlich SD. Insertional mutagenesis in Bacillus subtilis: mechanism and use in gene cloning. Gene. 1982 Oct;19(3):277–284. [PubMed]
  • Ohta H, Kato H, Okahashi N, Takahashi I, Hamada S, Koga T. Characterization of a cell-surface protein antigen of hydrophilic Streptococcus mutans strain GS-5. J Gen Microbiol. 1989 Apr;135(4):981–988. [PubMed]
  • Okahashi N, Sasakawa C, Okada N, Yamada M, Yoshikawa M, Tokuda M, Takahashi I, Koga T. Construction of NotI restriction map of the Streptococcus mutans genome. J Gen Microbiol. 1990 Nov;136(11):2217–2223. [PubMed]
  • OKAZAKI R, OKAZAKIT, STROMINGER JL, MICHELSON AM. Thymidine diphosphate 4-keto-6-deoxy-d-glucose, an intermediate in thymidine diphosphate L-rhamnose synthesis in Escherichia coli strains. J Biol Chem. 1962 Oct;237:3014–3026. [PubMed]
  • OUCHTERLONY O. Diffusion-in-gel methods for immunological analysis. Prog Allergy. 1958;5:1–78. [PubMed]
  • Perry D, Wondrack LM, Kuramitsu HK. Genetic transformation of putative cariogenic properties in Streptococcus mutans. Infect Immun. 1983 Aug;41(2):722–727. [PMC free article] [PubMed]
  • Pritchard DG, Furner RL. Structure of the group-specific polysaccharide of group E Streptococcus. Carbohydr Res. 1985 Dec 1;144(2):289–296. [PubMed]
  • Rajakumar K, Jost BH, Sasakawa C, Okada N, Yoshikawa M, Adler B. Nucleotide sequence of the rhamnose biosynthetic operon of Shigella flexneri 2a and role of lipopolysaccharide in virulence. J Bacteriol. 1994 Apr;176(8):2362–2373. [PMC free article] [PubMed]
  • RANTZ LA, RANDALL E. Use of autoclaved extracts of hemolytic streptococci for serological grouping. Stanford Med Bull. 1955 May;13(2):290–291. [PubMed]
  • Reeves P. Evolution of Salmonella O antigen variation by interspecific gene transfer on a large scale. Trends Genet. 1993 Jan;9(1):17–22. [PubMed]
  • Reeves PR, Hobbs M, Valvano MA, Skurnik M, Whitfield C, Coplin D, Kido N, Klena J, Maskell D, Raetz CR, et al. Bacterial polysaccharide synthesis and gene nomenclature. Trends Microbiol. 1996 Dec;4(12):495–503. [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. [PMC free article] [PubMed]
  • Severijnen AJ, van Kleef R, Hazenberg MP, van de Merwe JP. Chronic arthritis induced in rats by cell wall fragments of Eubacterium species from the human intestinal flora. Infect Immun. 1990 Feb;58(2):523–528. [PMC free article] [PubMed]
  • Shiroza T, Kuramitsu HK. Construction of a model secretion system for oral streptococci. Infect Immun. 1993 Sep;61(9):3745–3755. [PMC free article] [PubMed]
  • Soell M, Lett E, Holveck F, Schöller M, Wachsmann D, Klein JP. Activation of human monocytes by streptococcal rhamnose glucose polymers is mediated by CD14 antigen, and mannan binding protein inhibits TNF-alpha release. J Immunol. 1995 Jan 15;154(2):851–860. [PubMed]
  • Stinson MW, Nisengard RJ, Bergey EJ. Binding of streptococcal antigens to muscle tissue in vitro. Infect Immun. 1980 Feb;27(2):604–613. [PMC free article] [PubMed]
  • STROMINGER JL, PARK JT, THOMPSON RE. Composition of the cell wall of Staphylococcus aureus: its relation to the mechanism of action of penicillin. J Biol Chem. 1959 Dec;234:3263–3268. [PubMed]
  • Wang L, Romana LK, Reeves PR. Molecular analysis of a Salmonella enterica group E1 rfb gene cluster: O antigen and the genetic basis of the major polymorphism. Genetics. 1992 Mar;130(3):429–443. [PMC free article] [PubMed]
  • Yamashita Y, Takehara T, Kuramitsu HK. Molecular characterization of a STreptococcus mutans mutant altered in environmental stress responses. J Bacteriol. 1993 Oct;175(19):6220–6228. [PMC free article] [PubMed]
  • Yanisch-Perron C, Vieira J, Messing J. Improved M13 phage cloning vectors and host strains: nucleotide sequences of the M13mp18 and pUC19 vectors. Gene. 1985;33(1):103–119. [PubMed]
  • Zhang L, al-Hendy A, Toivanen P, Skurnik M. Genetic organization and sequence of the rfb gene cluster of Yersinia enterocolitica serotype O:3: similarities to the dTDP-L-rhamnose biosynthesis pathway of Salmonella and to the bacterial polysaccharide transport systems. Mol Microbiol. 1993 Jul;9(2):309–321. [PubMed]

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

Formats:

Related citations in PubMed

See reviews...See all...

Cited by other articles in PMC

See all...

Links

Recent Activity

Your browsing activity is empty.

Activity recording is turned off.

Turn recording back on

See more...