Molecular analysis of the 3,6-dideoxyhexose pathway genes of Yersinia pseudotuberculosis serogroup IIA

Kessler, Annette C.; Haase, Antje; Reeves, Peter R.

doi:10.1128/jb.175.5.1412-1422.1993

Cited by 63 publications

(41 citation statements)

References 47 publications

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“…The best-documented examples are found among serovars of S. enterica (43). There are also reports of high degrees of rfb homology in different species with shared Rfb functions, for example, S. enterica serovar Typhimurium and Yersinia pseudotuberculosis (23). The same is true of E. coli O7 and Shigella boydii type 12 (58), but these bacteria are sufficiently closely related to be considered a single species (44).…”

Section: Resultsmentioning

confidence: 76%

Relationships between rfb gene clusters required for biosynthesis of identical D-galactose-containing O antigens in Klebsiella pneumoniae serotype O1 and Serratia marcescens serotype O16

1995

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The lipopolysaccharide O antigens of Klebsiella pneumoniae serotype O1 and Serratia marcescens serotype O16 both contain a repeating unit disaccharide of [33)-␤-D-Galf-(133)-␣-D-Galp-(13]; the resulting polymer is known as D-galactan I. In K. pneumoniae serotype O1, the genes responsible for the synthesis of D-galactan I are found in the rfb gene cluster (rfb KpO1 ). We report here the cloning and analysis of the rfb cluster from S. marcescens serotype O16 (rfb SmO16 ). This is the first rfb gene cluster examined for the genus Serratia. Synthesis of D-galactan I is an rfe-dependent process for both K. pneumoniae serotype O1 and S. marcescens serotype O16. Hybridization experiments with probes derived from each of the six rfb KpO1 genes indicate that the cloned rfb SmO16 cluster contains homologous genes arranged in the same order. However, the degree of homology at the nucleotide sequence level was sufficiently low that hybridization was detected only under low-stringency conditions. rfbAB SmO16 genes were subcloned and shown to encode an ABC-2 (ATP-binding cassette) transporter which is functionally identical to the one encoded by the corresponding rfb genes from K. pneumoniae serotype O1. The amino acid sequences of the predicted RfbA and RfbB homologs showed identities of 75.7% (87.9% total similarity) and 78.0% (86.5% total similarity), respectively. The last gene of the rfb KpO1 cluster, rfbF KpO1 , encodes a bifunctional galactosyltransferase which initiates the formation of D-galactan I. RfbF KpO1 and RfbF SmO16 are 57.6% identical (with 71.1% total similarity), and both show similarity with RfpB, the galactosyltransferase involved in the synthesis of Shigella dysenteriae type I O-polysaccharide. The G؉C contents of the rfbAB genes from each organism are quite similar, and values are lower than those typical for the species. However, the G؉C content of rfbF SmO16 (47.6%) was much higher than that of rfbF KpO1 (37.3%), despite the fact that the average for each species (52 to 60%) falls within the same range.Lipopolysaccharide (LPS) is located in the outer leaflet of gram-negative bacteria and is responsible for the biological activity of endotoxin. In enteric bacteria, this molecule consists of three regions: lipid A, the core oligosaccharide, and the O polysaccharide. Lipid A forms the outer leaflet of the outer membrane bilayer. The core oligosaccharide region is attached to lipid A, and the O polysaccharide extends outward from the core region. O polysaccharides are composed of repeating oligosaccharide units with variable numbers of sugar residues. There is extensive structural diversity within O polysaccharides (22, 64). The composition, sequence, linkage, and substitution of the individual monomer residues within a repeating unit is characteristic for a given LPS and the parental bacterial strain. Thus, the O polysaccharide gives rise to epitopes (O antigens) which contribute to the immunological specificity of LPS.Klebsiella pneumoniae serotype O1 produces a high-molecular-weight smooth LPS (S-LPS)...

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Section: Resultsmentioning

confidence: 76%

Relationships between rfb gene clusters required for biosynthesis of identical D-galactose-containing O antigens in Klebsiella pneumoniae serotype O1 and Serratia marcescens serotype O16

1995

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“…Furthermore, although genetic information on dideoxyhexose formation in Yersinia spp. is currently lacking, the abrupt divergence of region C can also be observed upon comparison of Yersinia serogroups IIA (abequose [18]) and VA (this study).…”

Section: Discussionmentioning

confidence: 80%

“…Initiation towards this goal began with the cloning and analysis of the Salmonella typhimurium rfb (0-antigen) cluster, which led to the elucidation of the entire sequence of this region and the identification of 12 of the 15 genes present in the cluster (6,15,46). Subsequently, the deoxy and dideoxy genes within the rfb clusters of S. typhimurium (group B, which contains abequose) were compared to those within Salmonella paratyphi (group A, which contains paratose), Salmonella typhi (group D, which contains tyvelose), and, most recently, Y pseudotuberculosis M85 (serogroup IIA, which contains abequose) (17,18). As shown in Fig.…”

Section: Discussionmentioning

confidence: 99%

Studies of the biosynthesis of 3,6-dideoxyhexoses: molecular cloning and characterization of the asc (ascarylose) region from Yersinia pseudotuberculosis serogroup VA

et al. 1994

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The 3,6-dideoxyhexoses are found in the lipopolysaccharides of gram-negative bacteria, where they have been shown to be the dominant antigenic determinants. Of the five 3,6-dideoxyhexoses known to A large degree of bacterial immunological diversity in gram-negative species is attributed to the portion of the lipopolysaccharides known as the 0 antigen. Exposed on the cell envelope, the 0 antigen is the dominant surface entity which is polymorphic and provides the basis for the serological classification of the family Enterobacteriaceae (4,16,26). Among the large number of monosaccharides found as components of 0-specific polysaccharides, derivatives of the 3,6-dideoxyhexoses have drawn special attention because of their highly immunogenic characteristics (22,43,45). It has been shown that only a limited number of species of gram-negative bacteria, all in the family Enterobacteraceae, are able to produce these unusual sugars, and, of the eight possible stereoisomers of 3,6-dideoxyhexoses, only five have been identified in nature. Although colitose (3, and abequose (3,6-dideoxy-D-ylo-hexose, compound I [see Fig. 1]) can be detected in the lipopolysaccharide of some Escherichia coli serotypes and Citrobacter strains, respectively, the five 3,6-dideoxyhexoses known to occur naturally are present mainly in Salmonella and Yersinia spp. as major antigenic determinants (23). For example, abequose, colitose, tyvelose (3,6-dideoxy-D-arabino-hexose, compound II), and paratose (3,6-dideoxy-D-ribo-hexose, compound III) have been found in strains of Salmonella enterica (34), while all five, including ascarylose (3,6-dideOxy-L-arabino-hexose, compound * Corresponding author. Phone: (612) 626-7541. t Present address:

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“…RfbF protein of Salmonella typhimurium, which is 32.7% identical in 251 amino acids to Vig9 protein, catalyzes the synthesis of CDP-glucose from glucose 1-phosphate and CTP in the biosynthetic pathway of O-antigen (24). RfbF protein of Yersinia pseudotuberculosis is 31.1% identical in 257 amino acids to Vig9 protein (25). RfbA protein of Yersinia enterocolitica is 27.6% identical in 254 amino acids and catalyzes the synthesis of dTDP-L-rhamnose.…”

Section: Resultsmentioning

confidence: 99%

Saccharomyces cerevisiae VIG9 Encodes GDP-mannose Pyrophosphorylase, Which Is Essential for Protein Glycosylation

Hashimoto

Sakakibara

Yamasaki

et al. 1997

Journal of Biological Chemistry

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A genomic DNA fragment that complements a newly identified protein glycosylation-defective mutation, vig9, of Saccharomyces cerevisiae was cloned. Chromosomal integration of this fragment by homologous recombination indicated that it contains the wild type VIG9 gene. The nucleotide sequence was determined. A predicted gene product showed significant amino acid sequence homology with several bacterial enzymes that catalyze the synthesis of (deoxy)ribonucleotide diphosphate sugars from sugar phosphates and (deoxy)ribonucleotide triphosphate. We examined the enzyme activity to synthesize GDP-mannose in the cell extracts of the wild type, vig9 -1 mutant, and VIG9 transformant yeasts. Reduction of the activity in the mutant cell and its restoration by VIG9 suggested that the VIG9 gene is the structural gene for GDP-mannose pyrophosphorylase of S. cerevisiae which catalyzes the production of GDPmannose. We demonstrated the enzyme activity of Vig9 protein using a recombinant fusion protein produced in Escherichia coli.Most secretory and membrane proteins of eukaryotic cells are modified by glycosylation. Glycosylation endows protein with a wide variety of characteristics that are important in their cellular functions. Glycosylation occurs in the secretory pathway, and three types of modification are known: (i) attachment of N-linked saccharides to asparagine residues; (ii) attachment of O-linked saccharides to serine or threonine residues; and (iii) attachment of glycosylphosphatidylinositol anchors at the COOH termini. This is also the case with the yeast Saccharomyces cerevisiae, although mannose is the main component of N-and O-linked saccharides, and varieties of sugars are found in those of higher eukaryotes. In yeasts, the fourth type of protein-saccharide linkage was suggested for the covalent linkage between the cell wall proteins and glucans, although details are yet to be studied (1). Several excellent reviews on glycosylation in yeasts have been published (2)(3)(4)(5).N-Linked saccharides are first transferred to proteins as a Glc 3 Man 9 GlcNAc 2 unit from the Dol 1 -PP-intermediate when proteins are translocated into the endoplasmic reticulum. They are then trimmed to form the core N-linked saccharide, Man 8 GlcNAc 2 , which is common in eukaryotes. After proteins are transported from the endoplasmic reticulum to the Golgi apparatus in S. cerevisiae, core N-linked saccharides receive various quantities of mannose, depending on the protein species, one by one from GDP-Man. O-Linked saccharides that are composed exclusively of mannose in S. cerevisiae are also transferred to proteins in the Golgi apparatus sequentially from GDP-Man except for the first mannose, which is transferred from Man-P-Dol in the endoplasmic reticulum. As mannose is the major component of both N-and O-linked saccharides in S. cerevisiae, formation of GDP-Man, the activated form of mannose, should be vitally important.In S. cerevisiae, genetic approaches have been utilized in many scientific studies. To study the mechanism and function...

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Molecular analysis of the 3,6-dideoxyhexose pathway genes of Yersinia pseudotuberculosis serogroup IIA

Cited by 63 publications

References 47 publications

Relationships between rfb gene clusters required for biosynthesis of identical D-galactose-containing O antigens in Klebsiella pneumoniae serotype O1 and Serratia marcescens serotype O16

Relationships between rfb gene clusters required for biosynthesis of identical D-galactose-containing O antigens in Klebsiella pneumoniae serotype O1 and Serratia marcescens serotype O16

Studies of the biosynthesis of 3,6-dideoxyhexoses: molecular cloning and characterization of the asc (ascarylose) region from Yersinia pseudotuberculosis serogroup VA

Saccharomyces cerevisiae VIG9 Encodes GDP-mannose Pyrophosphorylase, Which Is Essential for Protein Glycosylation

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