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At least 19 recordsLinked to original sources
PubMed1978

The effect of some divalent cations on extracellular polysaccharide synthesis in Streptococcus salivarius.

The amount of extracellular insoluble polysaccharide produced by Streptococcus salivarius can be effected by some divalent cations. Calcium at concentrations of 1 X 10(-3) and 10(-4) M caused a reduction in polysaccharide synthesis. Magnesium at 1 X 10(-3) M inhibited extracellular polysaccharide production but at 1 X 10(-4) M had little effect. Manganese was without effect on polysaccharide synthesis. Zinc at 1 X 10(-3) and 1 X 10(-4) M caused a substantial increase in extracellular polysaccharide synthesis.

Calcium↗
PubMed1979

Outer membrane protein a and other polypeptides regulate capsular polysaccharide synthesis in E. coli K-12.

capR (lon) mutants of Escherichia coli K-12 are mucoid on minimal agar because they produce large quantities of capsular polysaccharide. When such mutants are transformed to tetracycline resistance by plasmid pMC44, a hybrid plasmid that contains a 2 megadalton (Mdal) endonuclease EcoR1 fragment of E. coli K-12 DNA joined to the cloning vehicle-pSC101, capsular polysaccharide synthesis is inhibited and the transformed colonies exhibit a non-mucoid phenotype. Re-cloning of the 2 Mdal EcoR1 fragment onto plasmid pHA105, a min-colE1 plasmid, yielded plasmid pFM100 which also inhibited capsular polysaccharide synthesis in the capR mutants. A comparison of the polypeptides specified by both plasmids pFM100 and pMC44 in minicells demonstrated that seven polypeptide bands were specified by the 2 MDal DNA, one of which was previously demonstrated to be outer membrane protein a; also known as 3b or M2 (40 kilodaltons, Kdal). Plasmid mutants no longer repressing capsular polysaccharide synthesis were either unable to specify the 40 Kdal outer membrane protein a or were deficient in synthesis of 25 Kdal and 14.5 Kdal polypeptides specified by the 2 Mdal DNA fragments. Studies with a minicell-producing strain that also contained a capR mutation indicated that the capR gene product regulated processing of at least one normal protein, the precursor of outer membrane protein a.

Bacterial Proteins↗
PubMed1977

Influence of salivary components and extracellular polysaccharide synthesis from sucrose on the attachment of Streptococcus mutans 6715 to hydroxyapatite surfaces.

The adsorption of (3)H-labeled Streptococcus mutans 6715 cells to disks of hydroxyapatite (HA) was studied. The number of streptococci that adsorbed was logarithmically related to the concentration of cells available up to at least 2 x 10(8) per ml; equilibrium occurred within 45 min. Assay reliability was verified by direct scanning electron microscopic counts. Untreated HA disks exposed to buffered saline (PBS)-suspended streptococci at a concentration of 1.1 x 10(8) per ml absorbed 3.2 x 10(6) cells per cm(2); approximately 3% of the surface area was, therefore, occupied by adsorbed organisms. The presence of adsorbed salivary components on HA reduced the number of attaching S. mutans cells by half. When S. mutans cells were suspended in saliva to mimic conditions existing in the mouth, the number of streptococci adsorbing to saliva-treated HA was reduced more than 30-fold compared to untreated HA. Approximately one-half of the streptococci adsorbed to untreated or to saliva-treated HA disks could be desorbed over a 4-h period with 0.067 M phosphate buffer. S. mutans cells exposed to sucrose to permit extracellular polysaccharide synthesis before or during adsorption attached in fewer numbers to both saliva-treated and untreated HA than PBS-treated organisms. When S. mutans cells adsorbed on untreated HA were exposed to sucrose, fewer organisms could be desorbed; thus, in situ polysaccharide synthesis promoted their more firm attachment to untreated HA. However, when saliva-suspended streptococci were adsorbed to saliva-treated HA surfaces, exposure to sucrose before or subsequent to adsorption did not promote more firm attachment. Evidently, the powerful adherence-inhibiting and desorptive effects of salivary components overshadowed any promoting effects attributable to glucan synthesis from sucrose. Similarly, no differences were noted in the desorption of S. mutans cells from human teeth after exposure to sucrose, glucose, or PBS relative to a strain of Streptococcus mitis (S. mitior). Thus, no evidence was obtained to support the hypothesis that glucan synthesis from sucrose was essential for, or promoted, the attachment of S. mutans cells to HA surfaces exposed to saliva or to the smooth surfaces of human teeth.

Adsorption↗
PubMed2025

Polysaccharide synthesis operon modulates Rickettsia-endothelial cell interactions.

Pathogenic Rickettsia species target vascular endothelial cells and cause systemic vasculitis. As obligate intracellular bacterial pathogens, Rickettsia must secure nutritional resources within the cytoplasm of endothelial cells while simultaneously subverting the innate immune defense system. With advances in rickettsial and host genetics, recent studies have identified novel molecular mechanisms involved in the complex interactions between Rickettsia and endothelial cells. However, it remains unclear how Rickettsia shields pathogen-derived immune stimulants, such as lipopolysaccharides (LPS) and peptidoglycan fragments, from immune recognition during intracellular replication. Prior work described two Rickettsia conorii variants with kkaebi transposon insertions in the polysaccharide synthesis operon (pso). Biochemical and immunological analyses revealed that pso is responsible for the biosynthesis of O-antigen (O-Ag) and the proper assembly of surface proteins. In the present work, we document that pso variant HK2 exhibits reduced capacities to adhere to and invade microvascular endothelial cells. Despite the low intracellular abundance, HK2 induced significantly higher levels of proinflammatory cytokines and chemokines, leading to premature cell death. Notably, HK2 exhibited defective intracellular survival in bone marrow-derived macrophages. This inability to dampen endothelial cell-mediated immune stimulation and resist macrophage-induced bactericidal activities resulted in the rapid elimination of viable Rickettsia in the mouse model of spotted fever. Further, when tested as a live-attenuated vaccine, HK2 elicited robust protective immunity against lethal spotted fever pathogenesis. Our work highlights the crucial role of pso in enabling Rickettsia to evade immune surveillance during intracellular replication within endothelial cells, ultimately delaying pathogen-induced programmed cell death and escaping immune defense mechanisms.

Operon↗
PubMed1975

Variation in internal polysaccharide synthesis among Streptococcus mutans strains.

Five strains, representative of Streptococcus mutans genetic group III antigenic group d, synthesized and degraded less intracellular polysaccharide (IPS) then 17 strains representative of other S. mutans groups. The strains that synthesize IPS degraded it rapidly. The production of acid in titratable amounts from endogenous IPS was usually complete within 1 h. IPS synthesis in S. mutans increased abruptly at culture glucose concentrations between 0.2 and 0.5% and was quantitated as both iodine-and glucose oxidase-positive material in cell hyrolysates. IPS degradation was measured by acid production in a pH-stat maintained at 7. The existence within group III d of a strain recently shown to be cariogenic in experimental animals suggest that IPS may not be a prerequisite for virulence in these cariogenic bacteria.

Acids↗
PubMed1979

Bacterial polysaccharide synthesis: an experiment for the dental biochemistry laboratory.

A simple experiment is described for demonstrating the formation of a high molecular-weight polysaccharide from sucrose by a bacterial enzyme. The polysaccharide confers high viscosity on solutions. Dental students are encouraged to consider this as a component of the system leading to the formation of dental plaque and the initiation of carious lesions.

Biochemistry↗
PubMed1975

Effect of dextranase on the extracellular polysaccharide synthesis of Streptococcus mutans; chemical and scanning electron microscopy studies.

A dextranase preparation (AD17) partially purified from a culture liquor of Spicaria violacea strain IFO 6120 significantly inhibited the formation of artifcial dental plaque on a steel wire or on an extracted tooth surface. Changes in the surface morphology of Streptococcus mutans cells due to AD17 action were studied using scanning electron microscopy. S. mutans cells grown in 5% sucrose-containing broth were coated with sticky amorphous capsule-like material, whereas cells grown in sucrose in the presence of AD17 or in glucose instead of sucrose did not synthesize such capsular material. AK17 degraded commercially available dextrans of molecular weight 7 X 1(04) and 2 X 10(6) to liberate glucose and various oligosaccharides, including isomaltose. On the other hand, AD17 hydrolyzed the extracellular polysaccharides (mainly glucan in nature) of some strains of S. mutans to a limited degree. Only 15 to 36% of the total polysaccharides were hydrolyzed by AD1M with little release of isomaltose. Prolonged incubation of the polysaccharides from S. mutans with AD17 did not release additional reducing sugars, which indicates that AD17 did not contain alpha-1,3-glucanase activity. These results suggest that glucosidic linkages which are susceptible to AD17 may play an important role in the adherence of S. mutans cells to smooth surfaces.

Dental Plaque↗
PubMed1975

Isolation and characterization of a bacteriophage polysaccharide.

A high molecular weight heteropolysaccharide, composed of glucose, glucuronic acid, N-acetylglucosamine, and mannose in an approximate molar ratio of 1:2:2:5, respectively, was isolated from phage K-2 and from the soluble fraction of phage-infected Aerobacter aerogenes lysates. Treatment of pure phage with 8 M urea at 4 degrees quantitatively solubilizes the bound polysaccharide and capsular polysaccharide (Yurewicz, E.C., Ghalambor, M.A., Duckworth, D.H., and Heath, E.C. (1971) J. Biol. Chem. 246, 5607-5616) with the release of only traces of other phage constituents; on this basis, it was concluded that the polysaccharide, like the the glycanohydrolase, is externally localized in the phage structure. Phage polysaccharide and glycanohydrolase fractionate similarly on ion exchange resins and gel electrophoresis in sodium dodecyl sulfate, but each may be purified to homogeneity by the procedures employed. The biosynthesis of the polysaccharide was shown to be uniquely dependent upon phage K-2 infection by: (a) absence of the polysaccharide in cells, the culture filtrate, or sonicated extracts of uninfected cells; (b) kinetics of polysaccharide synthesis following phage infection; and (c) isotopic double-labeling experiments that demonstrated the synthesis of polysaccharide only after initiation of phage replication in infected cells.

Acetylglucosamine↗
PubMed1976

Isolation of rough mutants of Klebsiella aerogenes and their synthesis of polysaccharides.

Two mutants which lacked both capsular and lipopolysaccharide O-antigen polysaccharides were isolated from Klebsiella aerogenes serotype 2 by phage selection; these were designated rough mutants. The polysaccharide fractions solubilized by partial acid hydrolysis of the lipopolysaccharide from both the wild type and mutants were chromatographed on Sephadex G-50. Analysis of the fractions obtained confirmed that the rough mutants lacked the galactan portion of the molecule, which is analogous to the Salmonella O-antigen polysaccharide. Membranes prepared from wild-type K. aerogenes, from a non-mucoid strain (lacking capsule only), and from one of the rough mutants were used in incubation mixtures to compare the biosynthesis of polysaccharides by these organisms. The incorporation of sugar nucleotides into both lipid intermediates and polymer was followed. Results show that the transferases were apparently present in all membranes, while the polymerases were absent in both the non-mucoid and rough mutants.

Klebsiella pneumoniae↗
PubMed1975

Comparison of polysaccharides produced by Myxococcus strains.

Exopolysaccharides were prepared from cultures of four Myxococcus strains grown on solid and in liquid media, and also from the fruiting bodies. Lipopolysaccharides could be extracted with aqueous phenol from the vegetative bacteria, but were absent from microcysts. Mannose and D-glucose were present in all the exopolysaccharides and three of the lipopolysaccharides examined. Other monosaccharides identified in the exopolysaccharides were D-galactose, N-acetylglucosamine and N-acetylgalactosamine. The composition of the lipopolysaccharides was more complex than that of the exopolysaccharides and, in addition to the neutral hexoses and amino sugars, rhamnose was identified in two preparations and ribose in another. No lipopolysaccharide preparations contained O-methyl xylose or heptose. The polysaccharides secreted by the bacillary forms grown on solid or in liquid media closely resembled the polysaccharides isolated from the fruiting bodies, in which they provided a matrix surrounding the microcysts. Each pair of polysaccharides contained the same monosaccharides, although in slightly different proportions. Differences were found in preparations from different strains. These results suggest that in the development cycle of the genus Myxococcus, considerable use is made of pre-existing enzyme systems to synthesize the precursors necessary for polysaccharide synthesis. Any specific difference between the polysaccharide produced by the bacilli and that surrounding the microcysts may lie in the fine structure, rather than in the individual components.

Acetylgalactosamine↗
PubMed1978

Cell-cell recognition in Saccharomyces cerevisiae: regulation of mating-specific adhesion.

Mating-specific adhesion between haploid yeast cells of opposite mating type (a and alpha) was studied by using a quantitative agar plate assay. Washed a and alpha cells that had not previously been exposed to their respective opposite mating type ("naive" cells) adhered relatively weakly. In water, only 5 to 10% of the a cells stuck tightly enough to alpha cells to give rise subsequently to diploid clones on the assay plates. Under optimum conditions (pH 6 to 7, at least 0.1 M Nacl or 0.01 M Mg(2+)), there was about 20% adhesion. Nevertheless, this weak binding defined a mating type-specific interaction because, even under optimum conditions, the homologous interactions (a with a and alpha with alpha) yielded only 3 to 5% cohesion. In contrast to these results, washed cells that had been preincubated in the cell-free culture medium of their opposite mating type ("preconditioned" cells) adhered quite strongly. The degree of adhesion between preconditioned cells (40 to 50%) was essentially unaffected by extremes of ionic strength, pH, and temperature and by the absence of divalent cation. This strong interaction was also mating type specific since cohesion between preconditioned cells of like mating type was only about 5%. The increase in agglutinability was obtained if only the a cells were preconditioned and could be induced by highly purified preparations of natural or synthetically prepared alpha-factor, an oligopeptide pheromone released by the alpha cells. The appearance of increased adhesiveness was blocked by an inhibitor of RNA synthesis and by an inhibitor of protein synthesis, but not by an inhibitor of polysaccharide synthesis. Adhesion between preconditioned cells could be inhibited by pretreatment with functionally univalent succinylated concanavalin A or with extracts from preconditioned cells of the opposite mating type. These results confirm in a quantitative manner that the recognition between conjugating cells of S. cerevisiae is a developmentally regulated event that is under the control of the mating type locus.

Adhesiveness↗
PubMed2025

The T2T genome assembly of watershield (Brasenia schreberi) unveils genomic insights into aquatic adaptation.

Watershield (Brasenia schreberi), belonging to Cabombaceae within the order Nymphaeales, represents one of the early-diverged angiosperm lineages. This perennial floating leaf freshwater aquatic plant features submerged juvenile leaves enveloped in a thick layer of transparent gelatinous mucilage, aiding in its resistance to aquatic stress. However, the evolutionary history of the mechanisms underlying its specific phenotype remains unclear. In this study, we present the telomere-to-telomere level genome of B. schreberi, unveiling that it underwent two rounds of whole-genome duplications (WGDs) and a recent whole-genome triplication, with the most ancient WGD being shared by Nymphaeaceae. WGD and dispersed duplication significantly contributed to the expansion of gene families, which are primarily associated with environmental adaptation. Additionally, we discovered that mature leaves primarily conduct photosynthesis and may transport nutrients to underwater juvenile leaves for polysaccharide synthesis. We also identified an ancestral broad expression pattern of ABC genes, and the similar expression of anthocyanin biosynthesis genes across all flower organs resulted in entirely purple flowers. Our findings deepen the understanding of the evolution of this specific aquatic plant phenotypes.

Genome, Plant↗
PubMed1978

Changes in the rate of synthesis of wall polysaccharides during the cell cycle of yeast.

Reevaluation and comparison of seemingly contradictory literature data on the mode of synthesis of wall polysaccharides during the cell cycle of Saccharomyces cerevisiae explained the source of discrepancies and demonstrated their general consonance in the following points: 1. The rate of synthesis of glucan and mannan is not constant and does not increase continuously throughout the entire cell cycle. 2. The rate of synthesis of both polysaccharides is considerably reduced at the time of cell division and in the prebudding phase.

Cell Cycle↗
PubMed1978

Experimental limb lengthening by epiphyseal distraction.

Kirshner wires were placed either side of the right distal femoral epiphysis and a constant tension device applied a distracting force across the plate in rabbits. Growth increase was measured between the wires and found to be about 150% greater than the concurrent normal growth between 2 control (undistracted) wires on the left; such growth increase can occur in the absence of fracturing. The forces required to do this were between 1/5 and 1/10 of those shown to cause fracturing in vitro. The growth increase was shown to be associated with hyperplasia and hypertrophy of the plate, as well as an increased rate of cell division and sulfated polysaccharide synthesis. This was in turn shown to be associated with an increase in new bone formation.

Animals↗