Search PubMed⌕ Search

Biomedical subjects

R P Burchard

Publications and source records attributed to R P Burchard.

At least 19 recordsLinked to original sources

Hydrophobicity, adhesion, and surface-exposed proteins of gliding bacteria.

The cell surface hydrophobicities of a variety of aquatic and terrestrial gliding bacteria were measured by an assay of bacterial adherence to hydrocarbons (BATH), hydrophobic interaction chromatography, and the salt aggregation test. The bacteria demonstrated a broad range of hydrophobicities. Results among the three hydrophobicity assays performed on very hydrophilic strains were quite consistent. Bacterial adhesion to glass did not correlate with any particular measure of surface hydrophobicity. Several adhesion-defective mutants of Cytophaga sp. strain U67 were found to be more hydrophilic than the wild type, particularly by the BATH assay and hydrophobic interaction chromatography. The very limited adhesion of these mutants correlated well with hydrophilicity as determined by the BATH assay. The hydrophobicities of several adhesion-competent revertants ranged between those of the wild type and the mutants. As measured by the BATH assay, starvation increased hydrophobicity of both the wild type and an adhesion-defective mutant. During filament fragmentation of Flexibacter sp. strain FS-1, marked changes in hydrophobicity and adhesion were accompanied by changes in the arrays of surface-exposed proteins as detected by an immobilized radioiodination procedure.

Journal Article↗

Adhesion and motility of gliding bacteria on substrata with different surface free energies.

The adhesion and motility of several aquatic and terrestrial gliding bacteria on slides differing in their critical surface energies have been examined. In general, adhesion was tenacious on low-critical surface energy (hydrophobic) surfaces and tenuous on hydrophilic surfaces. Gliding was inhibited on very hydrophobic substrata and skittish on very hydrophilic surfaces.

Bacteria↗

Effect of temperature shifts on gliding motility, adhesion, and fatty acid composition of Cytophaga sp. strain U67.

Gliding motility and flipping of 25 degrees C-adapted Cytophaga sp. strain U67 were inhibited when the bacteria were shifted to a less than or equal to 12 degrees C environment; motility was not blocked by a shift to 13 degrees C. Bacteria adapted to 4 degrees C were motile over the entire 4 to 25 degrees C temperature range tested. U67 adhesion to the substratum appeared to be unaffected by temperature shifts. Bacteria adapted to 4 degrees C had higher proportions of unsaturated and branched-chain fatty acids than did those grown at 25 degrees C. When 25 degrees C-adapted bacteria were subjected to a gradual temperature decline, the time of reappearance of gliding competence at 4 to 5 degrees C was correlated with these changes in fatty acid composition.

Acclimatization↗

Surface proteins of the gliding bacterium Cytophaga sp. strain U67 and its mutants defective in adhesion and motility.

Surface proteins of the gliding bacterium Cytophaga sp. strain U67 that make contact with glass substrata were radioiodinated, using a substratum-immobilized catalyst (Iodo-Gen). At least 15 polypeptides were iodinated, fewer than the number labeled by surface biotinylation of whole cells; these polypeptides define the set of possible candidates for the surface protein(s) that mediates gliding-associated substratum adhesion. The labeling of three adhesion-defective mutants exhibited two characteristic patterns of surface iodination which involved addition, loss, or alteration of several polypeptides of high molecular weight. An adhesion-competent revertant of mutant Adh3 and one of Adh2 exhibited the wild-type labeling pattern. Two other Adh2 revertants resembled their adhesion-defective parent. The labeling pattern of surface polypeptides of a nongliding but adhesive cell strain was similar to that of the wild type.

Bacterial Adhesion↗

Interference reflection microscopic study of sites of association between gliding bacteria and glass substrata.

Sites of close contact between gliding Cytophaga sp. strain U67 cells and glass were examined by interference reflection microscopy. Site patterns changed during translocation and moved relative to the substratum, in contrast to previous interference reflection microscopy observations of fibroblast and amoeboid motility. Sinistral rotation around the long axis of the cell was coupled with gliding, except when curved cells traversed curvilinear pathways. Close contact was temporary, since cells flipped up off the substratum on one pole, pivoted, or were displaced laterally in collisions. Other members of the order Cytophagales and Myxococcus sp. demonstrated similar patterns of close association with substrata.

Bacterial Adhesion↗

Trail following by gliding bacteria.

Slime trails, which are deposited on surfaces by gliding bacteria and which serve as preferential pathways for gliding motility, were tested for the species specificity of their support of movement. Among the pairs of bacteria tested, a variety of gliding bacteria and a flagellated bacterium moved along trails of unrelated species. Thus, the trails did not serve as pheromones. Rather, they may have guided gliding elasticotactically. Some biological implications of this finding are considered.

Bacteroidetes↗

Effect of cell length on gliding motility of Flexibacter.

Short Flexibacter FS-1 cells, generated during logarithmic growth in glucose-salts medium or by filament fragmentation during the transition from log to stationary phase in rich medium, are unable to glide. Motility returns when cells elongate. This strain also dissociates stable, short, nongliding variants.

Cytophagaceae↗

Intracellular, periodic structures in the gliding bacterium Myxococcus xanthus.

Electron microscopic observations of thin sections of Myxococcus xanthus vegetative cells revealed the presence of cytoplasmic bundles of 4- to 5-nm-diameter filaments running longtitudinally below the cell membrane and terminating in association with the envelope near one pole. Part of each bundle demonstrated a herringbone-like periodicity (approximately 12-nm spacing). This structure was observed in cells from shake cultures and in gliding cells fixed by several methods. It is proposed that the structure may be attached to the envelope near both poles in gliding cells and that the motive force for motility may be provided by its contraction and relaxation. In one of four nongliding mutants examined, the periodicity was indistinct or lacking. In this mutant another structure, comprised of linearly arrayed beads, was observed in association with the filamentous bundle. Another structure, characterized by major, transverse bands (approximately 34 nm apart), occurred in patches that may traverse the diameter of the wild-type cells in which the structure was observed.

Cell Wall↗

Pigmentation phenotype instability in Myxococcus xanthus.

Cells of Myxococcus xanthus FB2 produce tan or yellow colonies. Subcultures of tan colonies yielded tan and yellow colonies and subcultures of most yellow colonies yielded only yellow colonies. Strain FB2 variants in which the color type is more stable were obtained. Yellow cells were distinguishable from tan by the presence of pigment(s) with an absorption maximum at 379 nm. Fluctuation Test experiments and the presence of this pigment(s) in liquid cultures of FB2 indicated that tan phenotype cells spontaneously became or segregated yellow cells in liquid culture. The frequency of appearance of yellow cells was increased in low density cultures (less than 10(6)/ml). The increase cannot be explained by differences in growth rates of the two phenotypes. No evidence that cell-cell contact or culture medium constituents affect the appearance of the yellow phenotype was found. Ultraviolet irradiation of FB2 resulted in an increased proportion of cells producing yellow colonies among the survivors. Greater UV resistance of yellow cells and UV-induced conversion of tan to yellow accounts for this increase. Low level photoreactivation of viability and of the tan phenotype occurred. Incubation of FB2 in medium containing mitomycin C, nalidixic acid, phenethyl alcohol, or at 36.5 degrees C also resulted in conversion of tan to yellow cells.

Myxococcales↗

Phage and defective phage of strains of Myxococcus.

1. Phage-like particles were found in the supernatants of cultures of strains of Myxococcus xanthus, M. virescens and M. fulvus. The largest number of such particles was associated with M. virescens V2. Most of the particles were similar in morphology to the virulent Myxococcus phage, MX-1. 2. Several new phages were isolated from soil and animal droppings. A new phage was isolated from cultures of M. virescens V2. All resembled phage MX-1 in morphology and were related to phage MX-1 serologically. One of these phage, om, was characterized by fractionation of its proteins by SDS-polyacrylamide gel electrophoresis and by analysis of restriction fragments of its DNA. The very close relatedness with MX-1 was confirmed by these techniques. Phage om, was found to exist in a state of pseudolysogeny with strains of M. virescens and M. fulvus. 3. Two types of bacteriocin-like activity were found associated with Myxococcus strains. In one case, the activity was extracted from chloroformkilled or from sonicated cells. In the second case it was associated with extracellular material. Strains of Salmonella and Cytophaga were found to be good indicators for this latter activity. These strains were found to be killed by phage MX-1. 4. The significance of these data for origin of the phages of myxococci are discussed and it is proposed that MX-1 and the newly isolated phages may be virulent mutants of a family of lysogenic phages.

Bacteriocins↗

Mutants of Myxococcus xanthus insensitive to glycerol-induced myxospore formation.

Mutants of Myxococcus xanthus FBt unable to form myxospores in response to 0.5 M glycerol arise spontaneously with a frequency of 1--3 X 10(-5). These mutants are designated glc. Ultraviolet mutagenesis increases the frequency to a maximum of 7% of the survivors. The reversion frequency following ultraviolet irradiation of spontaneous glc mutants is less than 10(-3). Of four glc mutants examined, none form myxospores in response to the alternative inducers, ethylene glycol and dimethyl sulphoxide. One glc mutant is induced by 1.5 M glycerol; strain FBt responds to this glycerol concentration with low efficiency myxospore formation. Strain FBt and glc mutants all produce myxospores with low efficiency in response to phenyl ethanol. Of 117 glc mutants tested, 109 form fruiting bodies containing mature myxospores; thus, mutations to the glc phenotype do not normally block myxospore formation within the fruiting cycle of the organism.

Dimethyl Sulfoxide↗

Chloramphenicol resistance in Myxococcus xanthus.

Derivatives of Myxococcus xanthus FB(t) resistant to chloramphenicol (25 mug/ml) arose spontaneously with a frequency of approximately 10(-7). One of these organisms (FB(t)Cam(1) (r)) was characterized. FB(t)Cam(1) (r) showed a unique type of phenotypic instability. After transfer from medium containing chloramphenicol to medium lacking the drug, resistance was lost after approximately one generation. The loss resulted in a sharp drop in the total number of chloramphenicol-resistant organisms and was not due to segregation of chloramphenicol-susceptible organisms during growth. Cell-free extracts of strain FB(t)Cam(1) (r) converted chloramphenicol to acetyl chloramphenicols in a fashion implicating activity of chloramphenicol acetyltransferase. This activity was lost simultaneously with the loss of chloramphenicol resistance after removal of the drug from cultures. Organisms with a similar phenotype to FB(t)Cam(1) (r) could be produced at high frequencies when strain FB(t) was exposed to low concentrations of chloramphenicol (2 to 5 mug/ml), to 3-acetylchloramphenicol (25 mug/ml), or to 1,3-diacetylchloramphenicol (25 mug/ml). Since strain FB(t) is capable of deacetylating acetyl chloramphenicols, these effects are probably all due to low concentrations of chloramphenicol. In the presence of chloramphenicol, FB(t)Cam(1) (r) produced fruiting bodies and myxospores on fruiting agar; however, glycerol-induced myxospore formation was inhibited. In the absence of the antibiotic, chloramphenicol resistance was maintained by glycerol-induced myxospores.

Chloramphenicol↗

Myxospore induction in a nondispersed growing mutant of Myxococcus xanthus.

Myxococcus xanthus RB5, a rough-colony-forming, nondispersed growing mutant of strain FBt, forms macroscopic, multicellular masses of radially oriented cells in shake cultures. The cells appear to be held together by slime fibrils. Physical and enzymatic methods to disrupt the spheres were unsuccessful as were attempts to isolate dispersed growing mutants. During incubation of the spheres in starvation medium, the cells within convert to myxospores, indistinguishable from those formed in fruiting bodies. Myxospores were also induced in artifically constructed, dense masses of cells of a nonmotile strain.

Agar↗