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J Bacteriol. Jul 1995; 177(13): 3721–3727.
PMCID: PMC177088

Identification and characterization of a Bacteroides gene, csuF, which encodes an outer membrane protein that is essential for growth on chondroitin sulfate.


Bacteroides thetaiotaomicron can utilize a variety of polysaccharides, including charged mucopolysaccharides such as chondroitin sulfate (CS) and hyaluronic acid (HA). Since the enzymes (chondroitin lyases I and II) that catalyze the first step in breakdown of CS and HA are located in the periplasm, we had proposed that the first step in utilization of these polysaccharides was binding to one or more outer membrane proteins followed by translocation into the periplasm, but no such outer membrane proteins had been shown to play a role in CS or HA utilization. Previously we have isolated a transposon-generated mutant, CS4, which was unable to grow on CS or HA but retained the ability to grow on disaccharide components of CS. This phenotype suggested that the mutation in CS4 either blocked the transport of the mucopolysaccharides into the periplasmic space or blocked the depolymerization of the mucopolysaccharides into disaccharides. We have mapped the CS4 mutation to a single gene, csuF, which is capable of encoding a protein of 1,065 amino acids and contains a consensus signal sequence. Although CsuF had a predicted molecular weight and pI similar to those of chondroitin lyases, it did not show significant sequence similarity to the Bacteroides chondroitin lyase II, a Proteus chondroitin ABC lyase, or two hyaluronidases from Clostridium perfringens and Streptococcus pyogenes, nor was any CS-degrading enzyme activity associated with csuF expression in Bacteroides species or Escherichia coli. The deduced amino acid sequence of CsuF exhibited features suggestive of an outer membrane protein. We obtained antibodies to CsuF and demonstrated that the protein is located in the outer membrane. This is the first evidence that a nonenzymatic outer membrane protein is essential for utilization of CS and HA.

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Selected References

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  • Cheng Q, Hwa V, Salyers AA. A locus that contributes to colonization of the intestinal tract by Bacteroides thetaiotaomicron contains a single regulatory gene (chuR) that links two polysaccharide utilization pathways. J Bacteriol. 1992 Nov;174(22):7185–7193. [PMC free article] [PubMed]
  • Coulton JW, Mason P, Cameron DR, Carmel G, Jean R, Rode HN. Protein fusions of beta-galactosidase to the ferrichrome-iron receptor of Escherichia coli K-12. J Bacteriol. 1986 Jan;165(1):181–192. [PMC free article] [PubMed]
  • Dalbey RE, Von Heijne G. Signal peptidases in prokaryotes and eukaryotes--a new protease family. Trends Biochem Sci. 1992 Nov;17(11):474–478. [PubMed]
  • Davison J, Heusterspreute M, Chevalier N, Ha-Thi V, Brunel F. Vectors with restriction site banks. V. pJRD215, a wide-host-range cosmid vector with multiple cloning sites. Gene. 1987;51(2-3):275–280. [PubMed]
  • Feldhaus MJ, Hwa V, Cheng Q, Salyers AA. Use of an Escherichia coli beta-glucuronidase gene as a reporter gene for investigation of Bacteroides promoters. J Bacteriol. 1991 Jul;173(14):4540–4543. [PMC free article] [PubMed]
  • Glaser JH, Conrad HE. Chondroitin SO4 catabolism in chick embryo chondrocytes. J Biol Chem. 1979 Apr 10;254(7):2316–2325. [PubMed]
  • Goldberg MB, Boyko SA, Butterton JR, Stoebner JA, Payne SM, Calderwood SB. Characterization of a Vibrio cholerae virulence factor homologous to the family of TonB-dependent proteins. Mol Microbiol. 1992 Aug;6(16):2407–2418. [PubMed]
  • Guiney DG, Jr, Hasegawa P, Davis CE. Expression in Escherichia coli of cryptic tetracycline resistance genes from bacteroides R plasmids. Plasmid. 1984 May;11(3):248–252. [PubMed]
  • Guiney DG, Yakobson E. Location and nucleotide sequence of the transfer origin of the broad host range plasmid RK2. Proc Natl Acad Sci U S A. 1983 Jun;80(12):3595–3598. [PMC free article] [PubMed]
  • Guthrie EP, Salyers AA. Use of targeted insertional mutagenesis to determine whether chondroitin lyase II is essential for chondroitin sulfate utilization by Bacteroides thetaiotaomicron. J Bacteriol. 1986 Jun;166(3):966–971. [PMC free article] [PubMed]
  • Guthrie EP, Shoemaker NB, Salyers AA. Cloning and expression in Escherichia coli of a gene coding for a chondroitin lyase from Bacteroides thetaiotaomicron. J Bacteriol. 1985 Nov;164(2):510–515. [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]
  • Heller K, Kadner RJ. Nucleotide sequence of the gene for the vitamin B12 receptor protein in the outer membrane of Escherichia coli. J Bacteriol. 1985 Mar;161(3):904–908. [PMC free article] [PubMed]
  • Hynes WL, Ferretti JJ. Sequence analysis and expression in Escherichia coli of the hyaluronidase gene of Streptococcus pyogenes bacteriophage H4489A. Infect Immun. 1989 Feb;57(2):533–539. [PMC free article] [PubMed]
  • Kotarski SF, Linz J, Braun DM, Salyers AA. Analysis of outer membrane proteins which are associated with growth of Bacteroides thetaiotaomicron on chondroitin sulfate. J Bacteriol. 1985 Sep;163(3):1080–1086. [PMC free article] [PubMed]
  • Linn S, Chan T, Lipeski L, Salyers AA. Isolation and characterization of two chondroitin lyases from Bacteroides thetaiotaomicron. J Bacteriol. 1983 Nov;156(2):859–866. [PMC free article] [PubMed]
  • LOWRY OH, ROSEBROUGH NJ, FARR AL, RANDALL RJ. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed]
  • Lundrigan MD, Kadner RJ. Nucleotide sequence of the gene for the ferrienterochelin receptor FepA in Escherichia coli. Homology among outer membrane receptors that interact with TonB. J Biol Chem. 1986 Aug 15;261(23):10797–10801. [PubMed]
  • Moore WE, Holdeman LV. Human fecal flora: the normal flora of 20 Japanese-Hawaiians. Appl Microbiol. 1974 May;27(5):961–979. [PMC free article] [PubMed]
  • Nau CD, Konisky J. Evolutionary relationship between the TonB-dependent outer membrane transport proteins: nucleotide and amino acid sequences of the Escherichia coli colicin I receptor gene. J Bacteriol. 1989 Feb;171(2):1041–1047. [PMC free article] [PubMed]
  • Salyers AA, Guthrie EP. A deletion in the chromosome of Bacteroides thetaiotaomicron that abolishes production of chondroitinase II does not affect survival of the organism in gastrointestinal tracts of exgermfree mice. Appl Environ Microbiol. 1988 Aug;54(8):1964–1969. [PMC free article] [PubMed]
  • Salyers AA, Kotarski SF. Induction of chondroitin sulfate lyase activity in Bacteroides thetaiotaomicron. J Bacteriol. 1980 Aug;143(2):781–788. [PMC free article] [PubMed]
  • Salyers AA, Pajeau M, McCarthy RE. Importance of mucopolysaccharides as substrates for Bacteroides thetaiotaomicron growing in intestinal tracts of exgermfree mice. Appl Environ Microbiol. 1988 Aug;54(8):1970–1976. [PMC free article] [PubMed]
  • Salyers AA, O'Brien M. Cellular location of enzymes involved in chondroitin sulfate breakdown by Bacteroides thetaiotaomicron. J Bacteriol. 1980 Aug;143(2):772–780. [PMC free article] [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]
  • Sato N, Shimada M, Nakajima H, Oda H, Kimura S. Cloning and expression in Escherichia coli of the gene encoding the Proteus vulgaris chondroitin ABC lyase. Appl Microbiol Biotechnol. 1994 Mar;41(1):39–46. [PubMed]
  • Shoemaker NB, Getty C, Gardner JF, Salyers AA. Tn4351 transposes in Bacteroides spp. and mediates the integration of plasmid R751 into the Bacteroides chromosome. J Bacteriol. 1986 Mar;165(3):929–936. [PMC free article] [PubMed]
  • Shoemaker NB, Getty C, Guthrie EP, Salyers AA. Regions in Bacteroides plasmids pBFTM10 and pB8-51 that allow Escherichia coli-Bacteroides shuttle vectors to be mobilized by IncP plasmids and by a conjugative Bacteroides tetracycline resistance element. J Bacteriol. 1986 Jun;166(3):959–965. [PMC free article] [PubMed]
  • Struyvé M, Moons M, Tommassen J. Carboxy-terminal phenylalanine is essential for the correct assembly of a bacterial outer membrane protein. J Mol Biol. 1991 Mar 5;218(1):141–148. [PubMed]
  • Vercellotti JR, Salyers AA, Bullard WS, Wilkins D. Breakdown of mucin and plant polysaccharides in the human colon. Can J Biochem. 1977 Nov;55(11):1190–1196. [PubMed]

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