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Radiation response mechanisms of the extremely radioresistant bacterium Deinococcus radiodurans.

Effect of microgravity on recovery of bacterial cells from radiation damage was examined in IML-2, S/MM-4 and S/MM-9 experiments using the extremely radioresistant bacterium Deinococcus radiodurans. The cells were irradiated with gamma rays before the space flight and incubated on board the Space Shuttle. The survival of the wild type cells incubated in space increased compared with the ground controls, suggesting that the recovery of this bacterium from radiation damage was enhanced under the space environment. No difference was observed between the survivals of radiosensitive mutant rec30 cells incubated in space and on the ground. The amount of DNA-repair related RecA protein induced under microgravity was similar to those of ground controls, however, induction of PprA protein, product of a unique radiation-inducible gene (designated pprA) responsible for loss of radiation resistance in repair-deficient mutant, KH311, was enhanced under microgravity compared with ground controls. Recent investigation in vitro showed that PprA preferentially bound to double-stranded DNA carrying strand breaks, inhibited Escherichia coli exonuclease III activity, and stimulated the DNA end-joining reaction catalyzed by DNA ligases. These results suggest that D. radiodurans has a radiation-induced non-homologous end-joining (NHEJ) repair mechanism in which PprA plays a critical role.

Bacterial Proteins↗

[Method of algae-bacterium biofilm to improve the water quality in eutrophic waters].

The quality of eutrophic waters was improved by the algae-bacterium biofilm, which covered over the artificial aquatic mat made from a kind of mend macromolecule material. Two series of experiments show that using the artificial aquatic mat technique, algae bloom was inhibited markedly. At the eleventh day, the transparency was increased from 6 cm to 62 cm; and the concentrations of COD(Mn), TP, TN and NH4(+) -N were decreased to 92.89%, 49.25% , 94.97% and 70.15% respectively. The DO concentration also kept in certain satisfied level for using the algae-bacterium biofilm method. The suitable and economical density was about 8 - 16 bunchs/m(2) when the ratio of the artificial aquatic mat length to water depth was 0.7.

Bacteria↗

Gene cloning and expression of the catalase from the hydrogen peroxide-resistant bacterium Vibrio rumoiensis S-1 and its subcellular localization.

The catalase gene, vktA, of the hydrogen peroxide-resistant bacterium Vibrio rumoiensis S-1 has been isolated and sequenced. Plasmid pBSsa1 was obtained by genome library screening and it complemented a catalase-less mutant, Escherichia coli strain UM2, indicating that pBSsa1 contained the catalase gene (vktA). The vktA gene consisted of an open reading frame of 1530 bp encoding a 508 amino-acid protein with a calculated size of 57657.79 Da. The deduced amino acid sequence showed high homology with that of bacterial group III catalases. Investigation of the subcellular catalase localization using an immuno-electron microscopical technique revealed that a larger amount of VktA catalase is present in the periplasmic space than in the cytoplasm. The periplasmic space of this bacterium can therefore be regarded as a reservoir of VktA catalase in V. rumoiensis S-1.

Journal Article↗

[Characterization of enzymatic degradation of microcystins by a new isolated bacterium].

A strain of bacterium capable of biodegrading microcystin (MC) RR and MC-LR was isolated from the sediments of Dianchi Lake. It was demonstrated that the enzymes in cell-free extract of this bacterium were responsible for the degradation of MC-RR and MC-LR, and the dead-end products of MC-RR and MC-LR catalyzed by these enzymes were observed on HPLC chromatograms. Results show that the optimum pH for the activities of these enzymes was in the range from 6.0 to 8.0, however the metal ions of Cu2+, Mn2+ and Zn2+ had no apparent effects on the enzymatic degradation of MC-RR and MC-LR.

Bacteria↗

Amino acid sequence of cytochrome c' from the purple photosynthetic bacterium Rhodospirillum rubrum S1.

The amino acid sequence of cytochrome c' from the purple photosynthetic bacterium Rhodospirillum rubrum S1 has been determined and is consistent with homology to cytochrome c' from the nonphotosynthetic bacterium Alcaligenes sp. NCIB 11015. There is 29% identity in the chosen alignment of these two proteins. R. rubrum cytochrome c' is composed of a single peptide chain of 126 amino acid residues with a single heme covalently bound near the COOH terminus. There is no sequence similarity to mitochondrial cytochrome c, except at the heme binding site.

Amino Acid Sequence↗

Nucleotide sequence, expression, and properties of luciferase coded by lux genes from a terrestrial bacterium.

The lux genes required for expression of luminescence have been cloned from a terrestrial bacterium, Xenorhabdus luminescens, and the nucleotide sequences of the luxA and luxB genes coding for the alpha and beta subunits of luciferase determined. The lux gene organization was closely related to that of marine bacteria from the Vibrio genus with the luxD gene being located immediately upstream and the luxE downstream of the luciferase genes, luxAB. A high degree of homology (85% identity) was found between the amino acid sequences of the alpha subunits of X. luminescens luciferase and the luciferase from a marine bacterium, Vibrio harveyi, whereas the beta subunits of the two luciferases had only 60% identity in amino acid sequence. The similarity in the sequences of the alpha subunits of the two luciferases was also reflected in the substrate specificities and turnover rates with different fatty aldehydes supporting the proposal that the alpha subunit almost exclusively controls these properties. The luciferase from X. luminescens was shown to have a remarkably high thermal stability being stable at 45 degrees C (t 1/2 greater than 3 h) whereas V. harveyi luciferase was rapidly inactivated at this temperature (t 1/2 = 5 min). These results indicate that the X. luminescens lux system may be the bacterial bioluminescent system of choice for application in coupled luminescent assays and expression of lux genes in eukaryotic systems at higher temperatures.

Amino Acid Sequence↗

Distinct properties of Escherichia coli products of plant-type ribulose-1,5-bisphosphate carboxylase/oxygenase directed by two sets of genes from the photosynthetic bacterium Chromatium vinosum.

We have recently described the existence of two sets of genes encoding ribulose-1,5-bisphosphate carboxylase/oxygenase (Rbu-P2 carboxylase), rbcA-rbcB and rbcL-rbcS, in the photosynthetic purple sulfur bacterium Chromatium vinosum (Viale, A.M., Kobayashi, H., and Akazawa, T. (1989) J. Bacteriol. 171, 2391-2400). These genes were cloned in plasmid vectors, and their expression was studied in Escherichia coli. Expression of rbcA-rbcB in E. coli was obtained under the control of its own promoter. On the other hand, expression of rbcL-rbcS in this host was not observed unless these genes were cloned under the control of the tac promoter. Purified rbcA-rbcB and rbcL-rbcS products from E. coli consisted of large and small subunits in equimolar ratios. They also showed very close elution profiles to Rbu-P2 carboxylase isolated from C. vinosum in size-exclusion chromatography columns, thus suggesting hexadecameric (L8S8) structures. Vmax of Rbu-P2 carboxylase were very similar for both enzymes, but the Km values for CO2 and ribulose 1,5-bisphosphate showed some differences. Immunochemical and N-terminal amino acid sequence analyses of the large and small subunits encoded by rbcA-rbcB and rbcL-rbcS also differed, especially at the level of the small subunits. The comparisons described above as well as the analysis of C. vinosum crude extracts by anion-exchange chromatography indicated that Rbu-P2 carboxylase encoded by rbcA-rbcB was the only species detected in the photosynthetic bacterium.

Amino Acid Sequence↗

Characterization of a gram-positive bacterium from the proventriculus of budgerigars (Melopsittacus undulatus).

The cellular, cultural, and biochemical characteristics of eight isolates of a large gram-positive bacillus that are commonly observed as apparently normal flora in the proventriculus of budgerigars (Melopsittacus undulatus) were determined. The bacterium was highly pleomorphic and changed markedly in both diameter and length when subcultured on agar media. The bacterium was facultative anaerobic and capnophilic, hemolytic on blood agar, and formed flat colonies with irregular edges after incubation for several days. All isolates grew on sodium azide agar but did not grow on MacConkey agar. The isolates were catalase-negative and oxidase-negative and did not reduce nitrate. All isolates failed to utilize arginine, lysine, ornithine or tryptophane but produced acid from glucose, galactose, levulose, maltose, melibiose, starch, and sucrose. All isolates produced acetoin from glucose and hydrolyzed esculin. The eight isolates could not be identified to either genus or species level based on the descriptions of currently classified organisms in the division Firmicutes as described in Bergey's Manual of Systematic Bacteriology.

Animals↗

Purification and properties of a dicyclohexylcarbodiimide-sensitive adenosine triphosphatase from a thermophilic bacterium.

1. A stable ATPase complex with sensitivity to dicyclohexylcarbodiimide (TFo-F1) was purified from the membranes of the thermophilic aerobic bacterium PS3, by ion exchange chromatography in the presence of Triton X-100. 2. The ATPase of TFo-F1 was maximal at 70 degrees at pH 8.6 and was stable after monomerization in 4 M urea and 0.5% Triton X-100 at 25 degrees. The activity was dependent on Mg2+, Co2+, or Mn2+, and it became insensitive to dicyclohexylcarbodiimide when Ca2+ or Cd2+ was added instead. 3. TFo-F1 required P-lipids of this bacterium contained branched fatty acyl groups but no unsaturated groups and were stable against oxidation and heat. 4. Studies by electron microscopy, gel electrophoresis, and use of anti-ATPase antibody and [3H]acetyl-ATPase indicated that the TFo-F1 complex was composed of an ATPase moiety (TF1, five different subunits) and a hydrophobic moiety (TFo, three different subunits. TFo conferred TF1 with sensitivity to dicyclohexylcarbodiimide. 5. Vesicles catalyzing 32Pi-ATP exchange and ATP-driven enhancement of fluorescence of anilinonaphthalene sulfonate were reconstituted by dialyzing pure TFo-F1 and P-lipids together, and were active even at 50-75 degrees. The vesicles reconstituted from TFo-F1 and bacterial P-lipids were more stable than those reconstituted from TFo-F1 and soybean P-lipids.

Adenosine Triphosphatases↗

[Inheritance of hybrid plasmid pAS*-21 by cells of the purple phototropic nitrogen-fixing bacterium Rhodopseudomonas sphaeroides].

Clones of purple nitrogenfixing prototrophic bacterium Rhodopseudomonas sphaeroides carrying an incertion into the chromosome of the hybrid pAS8-121 (RP4-ColE1 (repA::Tn7] plasmid were analysed. It is revealed that plasmid integration could be due to both Tn7 and other migrating elements (IS8 and possibly, to resident migrating elements of purple bacteria). The plasmid pAS 8-121 can be autonomously transferred from the cointegrate state into Escherichia coli K-12; the plasmid is not inherited autonomously in cells of the purple bacterium. In E. coli cells R' derivatives of the plasmid carrying R. sphaeroides chromosomal fragments may be formed. The R' plasmids with fragments of plasmid DNA substituted for chromosomal material of R. sphaeroides were selected in E. coli K-12 cells among deleted (Kms) derivatives of pAS8-121.

Chromosomes, Bacterial↗

[Purification and properties of cytochrome c552 from purple sulfur bacterium Thiocapsa roseopersicina].

The method of purification up to electrophoretical homogeneity of cytochrome c552 from the phototrophic bacterium Thiocapsa roseopersicina, strain BBS is described. For the cytochrome absorption spectrum the maxima at 417, 523 and 552 nm are characteristic for the reduced state and at 409 nm--for the oxidized state. The molecular weight is equal to 62000. The cytochrome contains two hemes per molecule and consists of two subunits. pI is 4.1; E0' is about 10 mV. Cytochrome c552 is a flavoprotein according to its fluorescence spectrum and subunit structure. T. roseopersicina cytochrome c552 is able to be reduced with sulphide, cysteine and ascorbate as well as with H2 in the presence of hydrogenase from the same bacterium. These data suggest that cytochrome c552 from T. roseopersicina functions in vivo at the initial stage of electron transport from hydrogen and sulphide.

Chromatiaceae↗

Conjugation and meiosis of Paramecium caudatum infected with the micronucleus-specific bacterium Holospora elegans.

Cells of two complementary mating types of Paramecium caudatum, syngen 3, are infected with the micronucleus-specific bacterium Holospora elegans. Cells with bacteria in their micronuclei show the mating reaction with agglutination and pair formation. All the stages of meiosis in this species are observed and pronuclei are formed. During the pregamic divisions the bacteria are distributed to the division products, but many of the bacteria remain within the separation spindles, later being released into the medium. Therefore, the pronuclei contain only a few or no bacteria. In a test of the viability of exconjugant clones CNR-mutant (a behavioral mutant) and wild type cells, both infected with H. elegans, are crossed. The survival of exconjugants is very low (10.6% compared to 57.3% in the control). After conjugation only parental types are found indicating the occurrence of macronuclear regeneration after conjugation. The same result is obtained with cells which had contained bacteria but were cured by means of antibiotics. It is concluded that infected micronuclei have become genetically defective and did not give rise of new functional macronuclei. Therefore, H. elegans-bearing paramecia are genetically dead, which shows the parasitic nature of the bacterium H. elegans.

Animals↗

[ATP-dependent citrate lyase in the green phototrophic bacterium, Chlorobium limicola].

Cell-free extracts of the green sulfur bacterium Chlorobium limicola forma thiosulfatophilum, strains 1C and L, grown autotrophically and in media with organic substances were shown to cleave citrate with the formation of oxaloacetate and acetyl-CoA only when ATP, CoA and Mg2+ were added. These results suggest that C. limicola f. thiosulfatophilum produces ATP-linked citrate lyase (E. C. 4.1.3.8) which may play an important role in the carbon metabolism of this bacterium.

ATP Citrate (pro-S)-Lyase↗

Electron microscopy of a spiral-shaped bacterium in the blood and bone marrow of a rhinoceros iguana.

Spiral shaped bacteria present in blood smears and bone marrow of a sick rhinoceros iguana were examined by light and electron microscopy. The organisms averaged 10 micron in length and had at least three spiral turns. The cell contained nuclear areas, vacuoles and ribosomes, except at the poles where there was a virtual absence of organelles. The bacterium was found by a cell membrane, cell wall and enveloping sheath. "Blebs" were present with regularity on the cell surface. About 14 flagella were present at each pole, and at these sites there was a specialized thickening of the cell membrane. Organisms were present within a phagocytic vacuole of macrophages in the blood and bone marrow, and often these engulfed organisms were degenerated. The taxonomic position of the bacterium is unknown.

Animals↗

[Interaction in the phage-bacterium system].

The growth rate of the phage for Brevibacterium was studied as a function of the population of the host bacterium and its growth rate. The interaction was not additive when the phage-bacterium system was modeled in chemostat and in periodic regime. This fact was taken into account to describe more precisely and completely the qualitative characteristics of such a system as compared to models of the predator-victim type.

Bacteriophages↗

The 16S rRNA sequence and genome sizing of tributyltin resistant marine bacterium, strain M-1.

The 16S rRNA of the tributyltin resistant marine bacterium, strain M-1, was partly sequenced to confirm the taxonomic status. The results indicated that this bacterium should be classified under the genus Alteromonas, instead of a previous report in which this strain was identified as a Vibrio. The genome size of this strain was also measured by pulsed field gel electrophoresis (PFGE) using a contoureclamped homogeneous electric field. The strain was found to contain a genome size of 2,240 kilo base pairs, whereas Alteromonas nigrifaciens and Shewanella putrefaciens had 2,040 and 2,383 kilo base pairs, respectively. This is the first report of the genome sizing of the genus Alteromonas by PFGE.

Base Sequence↗

[Helicobacter pylori, a rediscovered bacterium. Implication in gastroduodenal diseases].

Helicobacter pylori is a microaerophilic bacterium initially found in the gastric antrum of patients with peptic ulcer disease. As a result, H. pylori is now believed to have a pathophysiologic role in gastritis as well as in peptic ulcer disease. Several recent studies showed that it may be associated with duodenal ulcer relapse and that eradication therapy using antibiotics may significantly decrease the ulcer recurrence rate in duodenal ulcer patients. Moreover, epidemiological studies suggest that it may increase the relative risk of carcinoma in the stomach and preliminary studies seem to indicate that some low-grade lymphoma in the stomach may regress after H. pylori eradication. Although the mechanisms by which H. pylori induces mucosal injury and/or neoplasm is not clearly understood, several modifications in gastric functions have been reported. The most specific way of detecting H. pylori in tissue is a combination of culture and histologic staining of mucosal biopsy specimens obtained by endoscopy. Rapid urease test, cytology and PCR procedures performed on biopsies may give rapid, sensitive and specific results. Breath test using 13C- or 14C-radiolabelled urea and serology tests are of particular importance when H. pylori diagnosis is needed via no invasive procedures. Helicobacter pylori is supposed to interact with G and D cells. Gastrin and somatostatin are synthetized and released from antral G and gastric D cells respectively. The gastric D cells are in close contact with either G and parietal cells. Gastrin stimulates gastric acid secretion and epithelial gastric cell proliferation (parietal and EC-L cells) while somatostatin inhibits these effects. Chronic gastritis is associated with fundic duodenal ulcer disease. In this situation, basal gastrin and meal- or bombesin-stimulated gastrin in the serum (especially gastrin G17) have been found to be higher in H. pylori positive than in negative patients. Moreover, gastrin decreases up to normal levels after eradication of H. pylori. The long term effect of a such hypergastrinemia is not so far established. The mechanism underlaying hormonal modification is poorly understood. Since no G/D cell ratio modification could be found after H. pylori eradication while the amount of somatostatin increases, one would suggest functional alteration of either G or D cells in the H. pylori-related chronic gastritis. The role of inflammatory mediators on the gastrin release and the processing of progastrin induced by the bacterium need further investigations.

Duodenal Diseases↗