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Tn5-mediated bleomycin resistance in Escherichia coli requires the expression of host genes.

The transposon Tn5 expresses a gene, ble, whose product increases the viability of Escherichia coli and also confers resistance to the DNA-cleaving antibiotic bleomycin and the DNA-alkylating agent ethylmethanesulphonate. We find that the Ble protein induces expression of an alkylation inducible gene, aidC, and that both the AidC gene product and DNA polymerase I are required for Ble to confer bleomycin resistance. These findings support models in which Ble enhances DNA repair and suggest that Tn5 confers a fitness advantage to the host bacterium by increasing the repair of spontaneous DNA lesions. Such co-operation between a transposon and its host suggests that Tn5 is a symbiotic rather than a selfish DNA element.

Bacterial Proteins↗

Mycetoma.

We have reviewed the outstanding facts about mycetoma, including the history, first reports in India, the different causal agents according to their geographic distribution, mycologic characteristics, pathology, microbiology, behavior, evolution, immunology, experimental inoculations, associated bacterial complications, and osseous lesions. Various drugs available for treatment are mentioned and recommended. Therapeutic results depend on the age of the disease, and, above all, the bony involvement. N. brasiliensis mycetoma is one of the most frequent species and causes the greatest number of mycetoma cases in America, especially in Mexico.

Africa↗

Genome streamlining in a cosmopolitan oceanic bacterium.

The SAR11 clade consists of very small, heterotrophic marine alpha-proteobacteria that are found throughout the oceans, where they account for about 25% of all microbial cells. Pelagibacter ubique, the first cultured member of this clade, has the smallest genome and encodes the smallest number of predicted open reading frames known for a free-living microorganism. In contrast to parasitic bacteria and archaea with small genomes, P. ubique has complete biosynthetic pathways for all 20 amino acids and all but a few cofactors. P. ubique has no pseudogenes, introns, transposons, extrachromosomal elements, or inteins; few paralogs; and the shortest intergenic spacers yet observed for any cell.

Alphaproteobacteria↗

Origin and evolution of the AmpC beta-lactamases of Citrobacter freundii.

To determine whether the widespread clinical use of beta-lactams has been selective for Citrobacter freundii-derived alleles of plasmid ampC genes, we generated a Bayesian consensus phylogeny of the published ampC sequences and compared the MICs of 16 beta-lactam antibiotics for Escherichia coli strains containing cloned copies of the C. freundii ampC alleles. We found that for the majority of compounds investigated, there has been essentially no increase in beta-lactam resistance conferred by those alleles. We also found that ampC alleles from the chromosomes of two beta-lactam-sensitive C. freundii strains isolated in the 1920s, before the clinical use of antibiotics, were as effective at providing beta-lactam resistance in E. coli as were the plasmid-borne alleles from beta-lactam-resistant clinical isolates. These results suggest that selection for increased resistance to beta-lactam antibiotics has not been a significant force directing the evolution of the C. freundii ampC alleles found in beta-lactam-resistant clinical isolates.

Bacterial Proteins↗

Invasin production by Yersinia pestis is abolished by insertion of an IS200-like element within the inv gene.

The two enteropathogens Yersinia pseudotuberculosis and Yersinia enterocolitica penetrate eukaryotic cells in vitro through invasin, a surface-exposed protein. In contrast, Yersinia pestis, the causative agent of plague, is unable to enter mammalian cell lines, although the inv gene is present on its chromosome. Although 99.3% identical to the inv gene of Y. pseudotuberculosis, the Y. pestis inv gene was disrupted in its central region by a 708-bp IS200-like element. Multiple copies of this insertion sequence element were found within the genome of the plague bacillus.

Adhesins, Bacterial↗

Cloning and characterization of multigenes encoding the immunodominant 30-kilodalton major outer membrane proteins of Ehrlichia canis and application of the recombinant protein for serodiagnosis.

A 30-kDa major outer membrane protein of Ehrlichia canis, the agent of canine ehrlichiosis, is the major antigen recognized by both naturally and experimentally infected dog sera. The protein cross-reacts with a serum against a recombinant 28-kDa protein (rP28), one of the outer membrane proteins of a gene (omp-1) family of Ehrlichia chaffeensis. Two DNA fragments of E. canis were amplified by PCR with two primer pairs based on the sequences of E. chaffeensis omp-1 genes, cloned, and sequenced. Each fragment contained a partial 30-kDa protein gene of E. canis. Genomic Southern blot analysis with the partial gene probes revealed the presence of multiple copies of these genes in the E. canis genome. Three copies of the entire gene (p30, p30-1, and p30a) were cloned and sequenced from the E. canis genomic DNA. The open reading frames of the two copies (p30 and p30-1) were tandemly arranged with an intergenic space. The three copies were similar but not identical and contained a semivariable region and three hypervariable regions in the protein molecules. The following genes homologous to three E. canis 30-kDa protein genes and the E. chaffeensis omp-1 family were identified in the closely related rickettsiae: wsp from Wolbachia sp. , p44 from the agent of human granulocytic ehrlichiosis, msp-2 and msp-4 from Anaplasma marginale, and map-1 from Cowdria ruminantium. Phylogenetic analysis among the three E. canis 30-kDa proteins and the major surface proteins of the rickettsiae revealed that these proteins are divided into four clusters and the two E. canis 30-kDa proteins are closely related but that the third 30-kDa protein is not. The p30 gene was expressed as a fusion protein, and the antibody to the recombinant protein (rP30) was raised in a mouse. The antibody reacted with rP30 and a 30-kDa protein of purified E. canis. Twenty-nine indirect fluorescent antibody (IFA)-positive dog plasma specimens strongly recognized the rP30 of E. canis. To evaluate whether the rP30 is a suitable antigen for serodiagnosis of canine ehrlichiosis, the immunoreactions between rP30 and the whole purified E. canis antigen were compared in the dot immunoblot assay. Dot reactions of both antigens with IFA-positive dog plasma specimens were clearly distinguishable by the naked eye from those with IFA-negative plasma specimens. By densitometry with a total of 42 IFA-positive and -negative plasma specimens, both antigens produced results similar in sensitivity and specificity. These findings suggest that the rP30 antigen provides a simple, consistent, and rapid serodiagnosis for canine ehrlichiosis. Cloning of multigenes encoding the 30-kDa major outer membrane proteins of E. canis will greatly facilitate understanding pathogenesis and immunologic study of canine ehrlichosis and provide a useful tool for phylogenetic analysis.

Amino Acid Sequence↗

Desiccation tolerance of prokaryotes.

The removal of cell-bound water through air drying and the addition of water to air-dried cells are forces that have played a pivotal role in the evolution of the prokaryotes. In bacterial cells that have been subjected to air drying, the evaporation of free cytoplasmic water (Vf) can be instantaneous, and an equilibrium between cell-bound water (Vb) and the environmental water (vapor) potential (psi wv) may be achieved rapidly. In the air-dried state some bacteria survive only for seconds whereas others can tolerate desiccation for thousands, perhaps millions, of years. The desiccated (anhydrobiotic) cell is characterized by its singular lack of water--with contents as low as 0.02 g of H2O g (dry weight)-1. At these levels the monolayer coverage by water of macromolecules, including DNA and proteins, is disturbed. As a consequence the mechanisms that confer desiccation tolerance upon air-dried bacteria are markedly different from those, such as the mechanism of preferential exclusion of compatible solutes, that preserve the integrity of salt-, osmotically, and freeze-thaw-stressed cells. Desiccation tolerance reflects a complex array of interactions at the structural, physiological, and molecular levels. Many of the mechanisms remain cryptic, but it is clear that they involve interactions, such as those between proteins and co-solvents, that derive from the unique properties of the water molecule. A water replacement hypothesis accounts for how the nonreducing disaccharides trehalose and sucrose preserve the integrity of membranes and proteins. Nevertheless, we have virtually no insight into the state of the cytoplasm of an air-dried cell. There is no evidence for any obvious adaptations of proteins that can counter the effects of air drying or for the occurrence of any proteins that provide a direct and a tangible contribution to cell stability. Among the prokaryotes that can exist as anhydrobiotic cells, the cyanobacteria have a marked capacity to do so. One form, Nostoc commune, encompasses a number of the features that appear to be critical to the withstanding of a long-term water deficit, including the elaboration of a conspicuous extracellular glycan, synthesis of abundant UV-absorbing pigments, and maintenance of protein stability and structural integrity. There are indications of a growing technology for air-dried cells and enzymes. Paradoxically, desiccation tolerance of bacteria has virtually been ignored for the past quarter century. The present review considers what is known, and what is not known, about desiccation, a phenomenon that impinges upon every facet of the distributions and activities of prokaryotic cells.

Adaptation, Physiological↗

Horizontal transfer of Shiga toxin and antibiotic resistance genes among Escherichia coli strains in house fly (Diptera: Muscidae) gut.

Whether the house fly, Musca domestica L., gut is a permissive environment for horizontal transfer of antibiotic resistance and virulence genes between strains of Escherichia coli is not known. House flies were immobilized and force fed suspensions of defined, donor strains of E. coli containing chloramphenicol resistance genes on a plasmid, or lysogenic, bacteriophage-born Shiga toxin gene stx1 (bacteriophage H-19B::Ap1). Recipient strains were E. coli lacking these mobile elements and genes but having rifampicin as a selectable marker. Plasmid transfer occurred at rates of 10(-2) per donor cell in the fly midgut and 10(-3) in the fly crop after 1 h of incubation postfeeding. Bacteriophage transfer rate was approximately 10(-6) per donor cell without induction, but induction with mitomycin C increased rates of transfer to 10(-2) per donor cell. These findings show that genes encoding antibiotic resistance or toxins will transfer horizontally among bacteria in the house fly gut via plasmid transfer or phage transduction. The house fly gut may provide a favorable environment for the evolution and emergence of pathogenic bacterial strains through acquisition of antibiotic resistance genes or virulence factors.

Animals↗

A model of structure and action of Sau3AI restriction endonuclease that comprises two MutH-like endonuclease domains within a single polypeptide.

Sau3AI is a type II endonuclease that cleaves GATC sequences, producing sticky ends with 4-nucleotide 5'-overhangs. Its activity is inhibited by cytosine C5-methylation within the target sequence. In the N-terminus, Sau3AI exhibits sequence similarity to the GATC-specific single-strand nicking endonuclease MutH implicated in mismatch repair (Ban and Yang, 1998). Sequence analysis of Sau3AI and its homologs reveals that Sau3AI possesses an additional MutH-like domain in the C-terminus. Structure prediction suggests that the C-terminal domain lacks the endonuclease active site but retains all putative DNA-binding elements. As an illustration of these findings, a model of quaternary structure of Sau3AI complexed with the target DNA is presented. These predictions have implications for evolution, structure and function of bacterial DNA repair enzymes and restriction endonucleases.

Amino Acid Sequence↗

Mycetoma: retrospective study of 13 cases in Tunisia.

Mycetomas are inflammatory pseudotumors of chronic and progressive evolution, of fungal (Eumycetoma), or bacterial (Actinomycetoma) origin. We conducted a retrospective study of patients who have been consulted for mycetoma over a period of 28 years. Thirteen cases of mycetomas were collected during the 28-year period, 10 patients were of rural origin. The lesions were localized on the foot in all instances. There was no male predominance. Histopathological examination confirmed mycetoma in all 13 cases and the determination of the infectious agent in 9 cases: 4 actinomycotic and 5 fungal. Microbiological examination identified the species in 7 cases. Bone involvement was found in 10 cases. Eumycetomas were treated by ketoconazole. The other cases were treated as actinomycetomas by antibiotics. The antibiotic treatment was associated with surgical excision in 4 cases and with amputation in 1 case. Mycetomas are rare in Tunisia, and only observed sporadically, for this reason the diagnosis is usually late, with severe functional, psychological, and socioeconomic consequences.

Adolescent↗

[Some aspects of antibiotic resistance of oligotrophic bacteria].

Antibiotic resistance spectra of a large group of oligotrophic and eutrophic bacteria from the open soil and aquatic ecosystems were studied. It was shown that sometimes antibiotic resistance of the oligotrophs was of plasmid nature. A possible transfer of plasmid antibiotic resistance from oligotrophs to pathogenic eutrophic soil and aquatic bacteria in the natural ecosystems is discussed. It was demonstrated that there was no insurmountable transcription/translation barriers between oligotrophic and eutrophic bacteria though the two bacterial groups are taxonomically and evolutionally very distant. A hypothesis was proposed that oligotrophic bacteria are likely to be one of the possible pools of plasmid antibiotic resistance for pathogenic microbes. Plasmid-free strains of some oligotrophic bacteria were selected. With the method of antibioticograms, taxonomic patterns for oligotrophic bacteria of the central group were developed.

Drug Resistance, Microbial↗

Antimicrobial resistance in the intensive care unit.

Antimicrobial resistance remains a major clinical problem in the intensive care unit despite the introduction of potent new antibiotics and the application of infection control measures. Impaired antibiotic permeability, antibiotic inactivation and alteration of antibiotic target sites, typically in concert, are mechanisms by which this resistance occurs. Resistance traits may be chromosomally mediated, in which case they may be expressed constitutively or inducibly, or may be mediated by plasmids and/or transposons that may confer resistance to multiple drugs and facilitate spread to other organisms. Bacteria in which clinically significant resistance has now become a potential problem include methicillin-resistant staphylococci, enterococci, a wide variety of Enterobacteriaceae, Pseudomonas aeruginosa, Pseudomonas cepacia, and Xanthomonas (Pseudomonas) maltophilia. Judicious limitation in the use of antimicrobials, especially in prophylactic regimens; use of antibiotics in appropriate doses; and, where possible, avoidance of drugs to which resistance has been shown to emerge rapidly in a specific clinical setting will help to minimize the evolution and spread of resistant bacterial isolates.

Cross Infection↗

[Light activation of NADH and NADPH].

Illumination of NADH and NADPH by UV-light in the absence of oxygen resulted in the reduction of ferredoxin or methyl-viologen to cation-radical and under prolonged illumination to dihydrodipyridyl. The reaction may by accompanied by triplet and singlet exitation of NADH. It was shown that hematoporphyrin in aqueous solution photosensitized the reaction of NADH oxidation by ferredoxin and methylviologen to the visible region of the spectrum. Under light excitation the redox potentials of NADH and NADPH were increased up to the level exceeding the potential of hydrogen electrode. Illumination of NADH and NADPH by UV-light in the presence of bacterial hydrogenase resulted in hydrogen evolution. The reaction of hydrogen evolution could be sensitised towards the visible region of the spectrum by chlorophyll or chloroplasts.

Catalysis↗

Radiometric estimation of the replication time of bacteria in culture: an objective and precise approach to quantitative microbiology.

In a recently developed, two-compartment liquid scintillation vial, the evolution of 14CO2 resulting from bacterial metabolism of uniformly labeled d-glucose was measured sensitively, cumulatively, and automatically in a liquid-scintillation counter. In each of eight species tested, a period of log-linear expansion of cumulative counting rate with time was observed. The exponential increase in cumulative counting rate was related to cell replication time by the integral of a first-order differential equation. Within a given species, the replication time measured by radioassay was found to be remarkably constant, unaffected by a fourfold variation in the activity of added labeled d-glucose, insensitive to the presence of carrier dextrose, and independent of the number of bacteria in the initial inoculum over a range of five orders of magnitude. These experiments demonstrate that the replication rate of an organism in culture is a highly reproducible characteristic that is susceptible to precise radiometric measurement in fundamental units of time under a variety of experimental conditions.

Bacteria↗

Kdp-like system in Salmonella typhimurium LT-2.

Salmonella typhimurium LT-2, as Escherichia coli K12, was able to grow in a potassium concentration-dependent manner, down to a very low concentration (< 5 microM). Its metabolic swelling also was [K+]-dependent. When the cells were subjected to hyperosmotic shock, this ion was uptaken rapidly, probably due to a K(+)-high affinity transport-system, similar to the E. coli Kdp system. The shrinkage in presence of 0.6 M NaCl, however, was more noticeable in S. typhimurium, which expressed a smaller level of intracellular K+ than E. coli. The genetic locus responsible for the ability of S. typhimurium to grow in low [K+], was mapped in nitrosoguanidine mutants and localized around min 18, close to the gal operon. This asseveration was confirmed by experiments of reversion, conjugation, and transduction. The mutants required considerably more [K+] to grow and to swell than the parental strain; in addition, below 1 mM [K+], they showed less internal [K+].

Adenosine Triphosphatases↗

On the evolution of arginases and related enzymes.

Sequence analysis of the arginase/agmatine ureohydrolase family, important enzymes in arginine/agmatine metabolism and the urea cycle, reveals the similarity of arginases to formiminoglutamate hydrolase (hutG) in Klebsiella aerogenes and to a previously unidentified open reading frame adjacent to the HMf locus of the archaebacterium Methanothermus fervidus. The gene structure and distribution of these homologous proteins across primary kingdoms suggest that this family is another example of a primordial enzyme possibly present in the universal common ancestor and that can be used as phylogenetic marker.

Amino Acid Sequence↗

Eukaryotic genes in Mycobacterium tuberculosis could have a role in pathogenesis and immunomodulation.

Acquisition of new genetic material through horizontal gene transfer has been an important feature in the evolution of many pathogenic bacteria. Here, we report the presence of 19 genes of eukaryotic origin in the genome of Mycobacterium tuberculosis, some of which are unique to the M. tuberculosis complex. These genes, having been retained in the genome through selective advantage, most probably have key functions in the organism and in mammalian tuberculosis. We explore the role these genes might have in manipulation of the host immune system by altering the balance of steroid hormones.

Amino Acid Sequence↗

An association between ATP binding cassette systems, genome sizes and lifestyles of bacteria.

Comparative bioinformatic analyses of ATP binding cassette (ABC) systems encoded in bacterial genomes have been undertaken in order to examine whether the range and distribution of these systems correlates with niches occupied by different organisms. In general, bacteria with larger genomes were found to encode more ABC systems than those with smaller genomes. Environmental bacteria, generally containing the largest genomes, showed the greatest number and diversity of ABC systems. Extracellular bacteria have larger genomes and show higher relative numbers of ABC transporters in comparison to intracellular bacteria. Similar results were obtained when comparing bacteria with different respiratory requirements since aerobic bacteria have larger genomes and also display greater numbers of ABC systems than anaerobes. These results suggest that the number of ABC systems encoded in bacterial genomes correlates with genome size and also with the physiological niche in which bacteria live. Furthermore, the distribution of the ABC systems into families indicates that the process of reductive evolution is responsible for retaining particular types of ABC systems as bacteria adapt to particular niches.

ATP-Binding Cassette Transporters↗