Cloning and Sequencing of the Genes forN-Acetylglucosamine Use That Construct Divergent Operons (nagE-nagAC) fromVibrio choleraeNon-O1

Yamano, Naoko; Oura, Noriyoshi; Wang, Jingyu; Fujishima, Shizu

doi:10.1271/bbb.61.1349

Cited by 16 publications

(10 citation statements)

References 6 publications

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“…PEPdependent phosphorylation and specific uptake of N-acetylglucosamine occur in the S. olivaceoviridis DNgcE mutant (which lacks a functional gene for the binding protein of the ABC transporter Ngc), but not in the DNgcE/DPtsC2 mutant. This finding is in accordance with the fact that PtsC2 shows 52%, 52%, 45% and 42% identity with the EIIC domains in NagE proteins (EIICBA or EIICB) from Klebsiella pneumoniae (Vogler and Lengeler 1989), E. coli (Rogers et al 1988), Vibrio cholerae (Yamano et al 1997) and V. furnissii (Bouma and Roseman 1996), respectively, and 46%, 44% and 45% identity with the EIIC domains (EIICB or EIICBA) in PtsG proteins from E. coli (Erni and Zanolari 1986), Staphylococcus carnosus (Christiansen and Hengstenberg 1996) and Bacillus subtilis (Zagorec and Postma 1992), respectively (Fig. 4).…”

Section: Discussionsupporting

confidence: 88%

“…slightly less than PtsC2 (see above)] to the EIIC domains in the NagE proteins from K. pneumoniae (Vogler and Lengeler 1989), E. coli (Rogers et al 1988), V. cholerae (Yamano et al 1997) and V. furnissii (Bouma and Roseman 1996), respectively. It shares 45%, 43% and 40% identity with the EIIC domains of PtsG from E. coli (Erni and Zanolari 1986), S. carnosus (Christiansen and Hengstenberg 1996) and B. subtilis (Zagorec and Postma 1992), respectively, and 40% identity with MalX from E. coli (Reidl and Boos 1991).…”

Section: Discussionmentioning

confidence: 96%

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Streptomyces olivaceoviridis possesses a phosphotransferase system that mediates specific, phosphoenolpyruvate-dependent uptake of N-acetylglucosamine

et al. 2002

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We recently described the ABC transporter Ngc (encoded by the ncgEFG operon) from Streptomyces olivaceoviridis, the first of its kind to be shown to transport N-acetylglucosamine and N,N'-diacetylchitobiose (chitobiose). A chromosomal mutant carrying a disruption of the ngcE gene, which encodes the sugar binding protein, was still able to transport N-acetylglucosamine. This phenotype can now be attributed to a functional phosphoenolpyruvate (PEP)-dependent phosphotransferase system (PTS). Two adjacent homologous genes, ptsC1 and ptsC2, were identified, and deduced to encode proteins which are 56% identical and can be predicted to contain eight transmembrane regions. PtsC1 (432 amino acids) and PtsC2 (403 residues) each correspond to a single EIIC domain; such domains are otherwise known only in several bacterial multidomain permeases for glucose/mannose or N-acetylglucosamine. The C-terminal sequences of PtsC1 and PtsC2 correspond to the motifs LKTPGREP and LPTRGRES, respectively. The ptsB gene located upstream of ptsC1 is predicted to encode a homologue of the EIIB domains usually found in bacterial multidomain permeases. Physiological and biochemical analyses of ngcE mutants carrying disruptive insertions in ptsC1 or ptsC2 or both revealed that, when grown on N-acetylglucosamine, the membrane component PtsC2, unlike PtsC1, mediates PEP-dependent, specific (K(m)=5 micro M) transport of N-acetylglucosamine, but not of other hexoses. Cross-complementation of membrane and cytoplasmic fractions from the various mutants led to the conclusion that S. olivaceoviridis also expresses the functional soluble components HPr, EI and EIIA of the PTS system. During growth on xylose, uptake of this pentose occurred if ptsC1 or ptsC2 was intact, but not in a mutant containing disrupted forms of both genes.

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Section: Discussionsupporting

confidence: 88%

Section: Discussionmentioning

confidence: 96%

Streptomyces olivaceoviridis possesses a phosphotransferase system that mediates specific, phosphoenolpyruvate-dependent uptake of N-acetylglucosamine

et al. 2002

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“…The predicted chitinases are diverse and include enzymes that are related to chitinases of bacteria (Fjoh_4175, Fjoh_4555, Fjoh_4757), animals (Fjoh_4560), and plants (Fjoh_2608). Other predicted enzymes that may be involved in chitin utilization include an N-acetylglucosamine kinase (Fjoh_4589) related to mouse NagK (34), an N-acetylglucosamine-6-phosphate deacetylase (Fjoh_3974) related to Vibrio cholerae NagA (98), and four glucosamine-6-phosphate isomerases/deaminases (Fjoh_2029, Fjoh_2381, Fjoh_4812, Fjoh_4557) related to E. coli NagB (79). NagK, NagA, and NagB are predicted to function in sequence to convert N-acetylglucosamine into fructose-6-phosphate for entry into the Embden-Meyerhof-Parnas pathway.…”

Section: Resultsmentioning

confidence: 99%

Novel Features of the Polysaccharide-Digesting Gliding Bacterium Flavobacterium johnsoniae as Revealed by Genome Sequence Analysis

McBride

Xie

Martens

et al. 2009

Appl Environ Microbiol

215

253

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The 6.10-Mb genome sequence of the aerobic chitin-digesting gliding bacterium Flavobacterium johnsoniae (phylum Bacteroidetes) is presented. F. johnsoniae is a model organism for studies of bacteroidete gliding motility, gene regulation, and biochemistry. The mechanism of F. johnsoniae gliding is novel, and genome analysis confirms that it does not involve well-studied motility organelles, such as flagella or type IV pili. The motility machinery is composed of Gld proteins in the cell envelope that are thought to comprise the "motor" and SprB, which is thought to function as a cell surface adhesin that is propelled by the motor. Analysis of the genome identified genes related to sprB that may encode alternative adhesins used for movement over different surfaces. Comparative genome analysis revealed that some of the gld and spr genes are found in nongliding bacteroidetes and may encode components of a novel protein secretion system. F. johnsoniae digests proteins, and 125 predicted peptidases were identified. F. johnsoniae also digests numerous polysaccharides, and 138 glycoside hydrolases, 9 polysaccharide lyases, and 17 carbohydrate esterases were predicted. The unexpected ability of F. johnsoniae to digest hemicelluloses, such as xylans, mannans, and xyloglucans, was predicted based on the genome analysis and confirmed experimentally. Numerous predicted cell surface proteins related to Bacteroides thetaiotaomicron SusC and SusD, which are likely involved in binding of oligosaccharides and transport across the outer membrane, were also identified. Genes required for synthesis of the novel outer membrane flexirubin pigments were identified by a combination of genome analysis and genetic experiments. Genes predicted to encode components of a multienzyme nonribosomal peptide synthetase were identified, as were novel aspects of gene regulation. The availability of techniques for genetic manipulation allows rapid exploration of the features identified for the polysaccharide-digesting gliding bacteroidete F. johnsoniae.

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“…For some pathogens, GlcNAc has been reported to be utilized more efficiently than glucose [36]. V. cholerae has an efficient system for GlcNAc catabolism which includes functional involvement of various enzymes encoded by nag genes ( nag A, B, C, D, E) [40]. The nag genes and also the chitinase genes including chiA2 are up-regulated in presence of GlcNAc [2].…”

Section: Discussionmentioning

confidence: 99%

The Vibrio cholerae Extracellular Chitinase ChiA2 Is Important for Survival and Pathogenesis in the Host Intestine

et al. 2014

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In aquatic environments, Vibrio cholerae colonizes mainly on the chitinous surface of copepods and utilizes chitin as the sole carbon and nitrogen source. Of the two extracellular chitinases essential for chitin utilization, the expression of chiA2 is maximally up-regulated in host intestine. Recent studies indicate that several bacterial chitinases may be involved in host pathogenesis. However, the role of V. cholerae chitinases in host infection is not yet known. In this study, we provide evidence to show that ChiA2 is important for V. cholerae survival in intestine as well as in pathogenesis. We demonstrate that ChiA2 de-glycosylates mucin and releases reducing sugars like GlcNAc and its oligomers. Deglycosylation of mucin corroborated with reduced uptake of alcian blue stain by ChiA2 treated mucin. Next, we show that V. cholerae could utilize mucin as a nutrient source. In comparison to the wild type strain, ΔchiA2 mutant was 60-fold less efficient in growth in mucin supplemented minimal media and was also ∼6-fold less competent to survive when grown in the presence of mucin-secreting human intestinal HT29 epithelial cells. Similar results were also obtained when the strains were infected in mice intestine. Infection with the ΔchiA2 mutant caused ∼50-fold less fluid accumulation in infant mice as well as in rabbit ileal loop compared to the wild type strain. To see if the difference in survival of the ΔchiA2 mutant and wild type V. cholerae was due to reduced adhesion of the mutant, we monitored binding of the strains on HT29 cells. The initial binding of the wild type and mutant strain was similar. Collectively these data suggest that ChiA2 secreted by V. cholerae in the intestine hydrolyzed intestinal mucin to release GlcNAc, and the released sugar is successfully utilized by V. cholerae for growth and survival in the host intestine.

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Cloning and Sequencing of the Genes forN-Acetylglucosamine Use That Construct Divergent Operons (nagE-nagAC) fromVibrio choleraeNon-O1

Cited by 16 publications

References 6 publications

Streptomyces olivaceoviridis possesses a phosphotransferase system that mediates specific, phosphoenolpyruvate-dependent uptake of N-acetylglucosamine

Streptomyces olivaceoviridis possesses a phosphotransferase system that mediates specific, phosphoenolpyruvate-dependent uptake of N-acetylglucosamine

Novel Features of the Polysaccharide-Digesting Gliding Bacterium Flavobacterium johnsoniae as Revealed by Genome Sequence Analysis

The Vibrio cholerae Extracellular Chitinase ChiA2 Is Important for Survival and Pathogenesis in the Host Intestine

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