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PubMed1988

Interaction between wild-type, mutant and revertant forms of the bacterium Streptococcus sanguis and the bacterium Actinobacillus actinomycetemcomitans in vitro and in the gnotobiotic rat.

In vitro, Streptococcus sanguis inhibits the growth of Actinobacillus actinomycetemcomitans, a presumed aetiological agent of localized juvenile periodontitis. When provided with glucose and good aeration, a growing culture of Strep. sanguis was found to produce hydrogen peroxide at concentrations in excess of the maximum LD50 reported for strains of A. actinomycetemcomitans. This concentration of hydrogen peroxide also inhibits the growth of the producer organism. A mutant of Strep. sanguis was isolated that lacked the ability to produce alpha-haemolysis on blood agar. This mutant had less than 3 per cent of its parent's level of pyruvate-oxidase activity, and made no detectable hydrogen peroxide. In vitro, the mutant had also lost the ability to inhibit the growth of A. actinomycetemcomitans. A spontaneous revertant, isolated by its ability to produce alpha-haemolysis, was found to have regained parental levels of pyruvate-oxidase activity and hydrogen-peroxide production and could inhibit the growth of A. actinomycetemcomitans in vitro. A gnotobiotic rat model was used to demonstrate that Strep. sanguis and A. actinomycetemcomitans interact in vivo and that this interaction depends on hydrogen-peroxide production by Strep. sanguis.

Actinobacillus↗
PubMed1983

Higher order structure in the 3'-minor domain of small subunit ribosomal RNAs from a gram negative bacterium, a gram positive bacterium and a eukaryote.

An experimental approach was used to determine and compare the highest order structure within the 150 to 200 nucleotides at the 3'-ends of the RNAs from the small ribosomal subunits of Escherichia coli, Bacillus stearothermophilus and Saccharomyces cerevisiae. Chemical reagents were employed to establish the degree of stacking and/or accessibility of each adenosine, guanosine and cytidine. The double helices were probed with a cobra venom ribonuclease from Naja naja oxiana, and the relatively unstructured and accessible sequences were localized with the single strand-specific ribonucleases A, T1, T2 and S1. The data enabled the various minimal secondary structural models, proposed for the 3'-regions of the E. coli and S. cerevisiae RNAs, to be critically examined, and to demonstrate that the main common features of these models are correct. The results also reveal the presence and position of additional higher order structure in the renatured free RNA. It can be concluded that a high level of conservation of higher order structure has occurred during the evolution of the gram negative and gram positive eubacteria and the eukaryote in both the double helical regions and the "unstructured" regions. Several unusual structural features were detected. Multiple G X A pairings in two of the putative helices, which are compatible with phylogenetic sequence comparisons, are strongly supported by the occurrence of cobra venom ribonuclease cuts adjacent to, and in one case between, these pairings. Evidence is also provided for the stacking of an A X A pair within a double helix of the yeast RNA. Other special structural features include adenosines bulged out from double helices; such nucleotides, which are hyper-reactive, have been implicated in protein recognition in 5 S ribosomal RNA. The 3'-terminal regions of the RNAs are particularly important for the functioning of the ribosome. They are involved in mRNA, tRNA and ribosomal factor binding. The results reveal that while the functionally important RNA sequences tend to be conserved, they are not always accessible in the free RNA; the pyrimidine-rich "Shine and Dalgarno" sequence, for example, which is involved in mRNA recognition, occurs in a double helix in both eubacterial RNAs.

Base Sequence↗
PubMed1996

Crystalline alcohol dehydrogenases from the mesophilic bacterium Clostridium beijerinckii and the thermophilic bacterium Thermoanaerobium brockii: preparation, characterization and molecular symmetry.

Two tetrameric NADP(+)-dependent bacterial secondary alcohol dehydrogenases have been crystallized in the apo- and the holo-enzyme forms. Crystals of the holo-enzyme from the mesophilic Clostridium beijerinckii (NCBAD) belong to space group P2(1)2(1)2(1) with unit-cell dimensions a = 90.5, b = 127.9, c = 151.4 A. Crystals of the apo-enzyme (CBAD) belong to the same space group with unit-cell dimensions a = 80.4, b = 102.3, c = 193.5 A. Crystals of the holo-enzyme from the thermophilic Thermoanaerobium brockii (NTBAD) belong to space group P6(1(5)) (a = b = 80.6, c = 400.7 A). Crystals of the apo-form of TBAD (point mutant GI98D) belong to space group P2(1) with cell dimensions a = 123.0, b = 84.8, c = 160.4 A beta = 99.5 degrees. Crystals of CBAD, NCBAD and NTBAD contain one tetramer per asymmetric unit. They diffract to 2.0 A resolution at liquid nitrogen temperature. Crystals of TBAD(GI98D) have two tetramers per asymmetric unit and diffract to 2.7 A at 276 K. Self-rotation analysis shows that both enzymes are tetramers of 222 symmetry.

Journal Article↗
PubMed1995

Gene transfer from a bacterium injected into an aquifer to an indigenous bacterium.

Two novel 3-chlorobenzoate-degrading bacteria were previously isolated from an aquifer in which no such bacteria could be enriched prior to the introduction of the 3-chlorobenzoate-degrading strain, Pseudomonas sp. B13. To understand the origin of 3-chlorobenzoate-degrading genes in the two novel isolates, the 16S ribosomal RNA, clcD (dienelactone hydrolase) and clcA (chlorocatechol oxygenase) genes from these bacteria were amplified and sequenced. The partial 16S rRNA gene sequences and REP-PCR patterns showed that these two novel isolates were identical but differed from strain B13. Phylogenetic analyses revealed that the novel isolates were closely related to Alcaligenes eutrophus in the beta subclass of the Proteobacteria, whereas strain B13 was related to Pseudomonas aeruginosa and P. mendocina in the gamma subclass of the Proteobacteria. In contrast, the clcD and clcA gene sequences were identical on strain B13 and these two isolates, indicating that the 3-chlorobenzoate-degrading genes were transferred from strain B13 to these isolates. What cannot be established is when this transfer occurred.

Base Sequence↗
PubMed1988

Heat Production by the Denitrifying Bacterium Pseudomonas fluorescens and the Dissimilatory Ammonium-Producing Bacterium Pseudomonas putrefaciens during Anaerobic Growth with Nitrate as the Electron Acceptor.

The heat production rate and the simultaneous nitrate consumption and production and consumption of nitrite and nitrous oxide were monitored during the anaerobic growth of two types of dissimilatory nitrate reducers. Pseudomonas fluorescens, a denitrifier, consumed nitrate and accumulated small amounts of nitrite or nitrous oxide. The heat production rate increased steadily during the course of nitrate consumption and decreased rapidly concomitant with the depletion of the electron acceptors. A mean experimental enthalpy change value of -800 kJ/mol of nitrate and a mean growth yield value of 33 g (dry weight)/mol of nitrate consumed were obtained for different concentrations of nitrate. For Pseudomonas putrefaciens, a dissimilatory ammonium producer, the nitrate consumption resulted in an accumulation of nitrite and nitrous oxide. Nitrite consumption commenced after depletion of the nitrate; consequently, two phases were noted in the heat production rate curve during growth. A mean experimental enthalpy change value of -810 kJ/mol of nitrate was obtained for different concentrations of nitrate.

Journal Article↗
PubMed1998

A gliding bacterium strain inhibits adhesion and motility of another gliding bacterium strain in a marine biofilm

Two species of gliding bacteria were isolated from a marine biofilm. They were described and identified as members of the genus Cytophaga. One of them (RB1057) produced an extracellular inhibitor of colony expansion of the other (RB1058). The inhibitor was characterized as a glycoprotein with an apparent molecular mass of 60 kDa. It inhibited RB1058 adhesion to and gliding on substrata. Motility and adhesion of several other aquatic gliding bacteria were not measurably affected by this agent.

Journal Article↗