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Effect of ozone on prophage induction in different strains of Escherichia coli K-12 lysogenic for lambda.

Ozone was tested for its effect upon induction of lambda prophage in two different strains of Escherichia coli K-12. Based on the induction index and when compared to ultraviolet light, ozone appeared to be a weak, if any at all, inducer of the lytic cycle in E. coli. This is in agreement with other studies which have suggested that this agent is a weak inducer of the SOS functions.

Bacteriophage lambda↗

Prophage-mediated production of a bacteriocinlike substance by SP beta lysogens of Bacillus subtilis.

Cultures of Bacillus subtilis lysogenic for the temperature bacteriophage SP beta release "betacin", a bacteriocinlike substance that inhibits B. subtilis strains which do not carry this phage. Production of betacin is blocked by mutations in the bet gene on the prophage and a second phage gene, tol, is apparently involved in making the lysogen itself tolerant to betacin. Mutations in a bacterial gene betR, located on the B. subtilis chromosome between metC and pyrD, render nonlysogens tolerant to betacin.

Antigens, Bacterial↗

Complete nucleotide sequence of the prophage VT2-Sakai carrying the verotoxin 2 genes of the enterohemorrhagic Escherichia coli O157:H7 derived from the Sakai outbreak.

The enterohemorrhagic Escherichia coli (EHEC) O157:H7 strain RIMD 0509952, derived from an outbreak in Sakai city, Japan, in 1996, produces two kinds of verotoxins, VT1 and VT2, encoded by the stx1 and stx2 genes. In the EHEC strains, as well as in other VT-producing E. coli strains, the toxins are encoded by lysogenic bacteriophages. The EHEC O157:H7 strain RIMD 0509952 did not produce plaque-forming phage particles upon inducing treatments. We have determined the complete nucleotide sequence of a prophage, VT2-Sakai, carrying the stx2A and stx2B genes on the chromosome, and presumed the putative functions of the encoded proteins and the cis-acting DNA elements based on sequence homology data. To our surprise, the sequences in the regions of VT2-Sakai corresponding to the early gene regulators and replication proteins, and the DNA sequences recognized by the regulators share very limited homology to those of the VT2-encoding 933W phage carried by the EHEC O157:H7 strain EDL933 reported by Plunkett et al. (J. Bacteriol., p1767-1778, 181, 1999), although the sequences corresponding to the structural components are almost identical. These data suggest that these two phages were derived from a common ancestral phage and that either or both of them underwent multiple genetic rearrangements. An IS629 insertion was found downstream of the stx2B gene and upstream of the lysis gene S, and this might be responsible for the absence of plaque-forming activity in the lysate obtained after inducing treatments.

Bacterial Toxins↗

Prophage, phiPV83-pro, carrying panton-valentine leukocidin genes, on the Staphylococcus aureus P83 chromosome: comparative analysis of the genome structures of phiPV83-pro, phiPVL, phi11, and other phages.

Staphylococcus aureus P83 has Panton-Valentine leukocidin (PVL)-like genes, lukM and lukF-PV. Here, lukM and lukF-PV genes were found on the genome of a prophage, which was designated as phiPV83-pro. The precise genome size was 45,636 bp with att core sequences of 10 base pairs. Sixty-four ORFs were identified on the phiPV83-pro genome, including two extra operons, lukM-lukF-PV and orfs63-64. The lukM-lukF-PV cluster was located 2.1 kb upstream of the attL site. The most striking feature of the phiPV83-pro genome was a constituent of at least 4 regions from phi11, phiPVL, and other phages, i.e., (i) att sites identical with those of phi11, (ii) a cos sequence and the genes encoding packaging and head proteins of phiPVL (occupied half region of phiPV83-pro), and (iii) the other two regions which showed no significant similarity with known phages (occupied about 40% of phiPV83-pro). Furthermore, two insertion sequences, ISSA1 and ISSA2 were integrated into attL site and orf44, respectively. PhiPV83-pro was not induced as phage particles from S. aureus P83 regardless of its treatment with mitomycin C. The insertion of ISSA1 into the attL site was one of the reasons of the failure of the induction of the phage particles by mitomycin C treatment of the strain P83.

Amino Acid Sequence↗

Association of plasmid integrative J7W-1 prophage with Bacillus thuringiensis strains.

A region homologous to the genome of plasmid integrative phage J7W-1 was detected in the largest plasmid in 3 out of 22 type strains of Bacillus thuringiensis, dendrolimus (DEN), aizawai (AIZ) and indiana (IND). Phage induction by ethidium bromide observed particularly in the J7W-1 lysogen was identified in DEN and IND but not AIZ strains. The morphology of the phage induced in DEN and IND strains was identical to J7W-1, but the phage production in IND strain was lower as compared to the J7W-1 lysogen. Although the restriction analysis indicated that the prophage in DEN strain possessed a complete J7W-1 genome, modification and/or deletion had presumably occurred in AIZ and IND strains.

Bacillus Phages↗

Prophage induction and cell division in E. coli. III. Mutations sfiA and sfiB restore division in tif and lon strains and permit the expression of mutator properties of tif.

In E. coli K12, cell filamentation promoted by tif is enhanced by the lon mutation; in contrast, prophage induction and repair of UV-irradiated phage lambda, also promoted by tif, are not affected by lon. From a tif lon double mutant, "revertants" having recovered the ability to divide at 41 degrees were isolated, among which most (95%) had also lost their Lon filamentous phenotype after ultraviolet (UV) irradiation. From these 95% of revertants: (1) 94% are suppressed for the whole Tif phenotype, by additional mutations that render them deficient in DNA repair, as judged from their high UV sensitivity; some have been characterized as recA mutants. (2) 1% have recovered a control on cell division at 41 degrees or after UV irradiation by means of secondary mutations altering neither the other phenotypic properties of tif and lon, nor the repair and recombination ability of the cells: in particular, this class of "revertants" remains thermoinducible upon lysogenisation; the mutations which specifically suppress filamentation have been mapped at two loci, sfiA and sfiB, cotransducible respectively with pyrD and leu. In the remaining 5% of revertants that still exhibit an UV-induced filamentous growth, 3% can be tentatively classified as true tif+ revertants; 2% behave as tif thermodependent revertants, showing suppression of the Tif (and Lon) phenotype only at 41 degrees: 2recAts have been identified in this class. Non-lysogenic tif lon sfi and tif sfi strains remain viable during prolonged growth at 41 degrees. Under these conditions, tif expresses mutator properties, which can be conveniently analyzed in this sfi background. The action of lif, lon and sfi mutations is tentatively interpreted on the basis of a negative control of cell division specifically associated with DNA repair.

Cell Division↗

Genomic relatedness of Staphylococcus aureus phages of the International Typing Set and detection of serogroup A, B, and F prophages in lysogenic strains.

On the basis of HindIII-restriction digest analysis of genomic DNAs, the S. aureus bacteriophages of the International Typing Set were divided into five clusters designated as A, F, Ba, Bb, and Bc. The clusters A and F include all the phages of serogroups A and F and correspond to species 3A and 77 proposed by Ackermann and DuBow (1987). On the other hand, the phages of serogroup B were divided into three clusters designated as Ba, Bb, and Bc that differ significantly each from the other in their restriction patterns. The clusters Ba and Bb may represent two separate species, while the cluster Bc may include more than one phage species. For each of the phage serogroups A, B, and F, common HindIII-restriction fragments of phage 3A (1700 bp), of 53 (4060 bp), and of 77 (8300 bp) were used for the preparation of probes specific to the phages of serogroups A, B, and F. These probes were very effective, making it possible to detect up to three different prophages in a given lysogenic strain at the same time. Restriction enzyme maps of phages 3A, 53, and 77, each representing a different serogroup, were constructed. The restriction maps of phage 3A and that of phage 77 are linear, whereas that of phage 53 is circular and exhibits a circular permutation. DNAs of the phages of serogroups A and F have cohesive ends. On each restriction map, the sites corresponding to specific probes are indicated. The size of intact genomic DNA of all phages estimated by PFGE varies within the range of 41.5-46.2 kb.

Bacteriophage Typing↗

Mating in Bacillus thuringiensis can induce plasmid integrative prophage J7W-1.

Bacillus thuringiensis serovar israelensis, a bacterium which possesses plasmid transfer ability after mating, has been lysogenized by plasmid integrative phage J7W-1. The induction of phage in this J7W-1 lysogen was observed after mating with phage-insensitive strains, such as B. thuringiensis serovar thuringiensis, B. cereus and B. subtilis, as well as the phage-sensitive strain serovar israelensis. The phage induction was not observed after mating with B. thuringiensis strains AF101, serovar dendrolimus and serovar indiana. Because these strains are naturally associated with J7W-1 or its related phage, the data strongly suggest a constitutive expression of the repressor encoded by the prophage in these strains. However, the phage induction was observed in B. thuringiensis serovar aizawai, although it contained the J7W-1 DNA homologous region(s).

Bacillus Phages↗

[Structure of a region in the genome of bacteriophage N15, necessary for formation of hairpins at ends of the linear plasmid prophage].

A fragment containing telRL site of bacteriophage N15 has been cloned in the vector plasmid pUC19. The nucleotide sequence of a small region from EcoRV-PstI fragment has been defined by Maxam-Gilbert technique. The analysis of the obtained sequence has shown the telRL site to be a nonideal palindrome (the size of 56 nucleotide ops) in which two nucleotide pairs differ in the positions 12 and 14 on both sides of the palindrome centre. The DNA region with alteration of purines and pyrimidines (GC) surrounded by AT-rich regions: 5'-ATTATACGCGCGTATAAT-3'--in the symmetry centre of palindrome is characteristic of the telRL site structure. This characteristic of the region may play a key role in recognition of the site by the specific enzyme at formation of linear prophage-plasmid during lysogenization.

Bacteriophages↗

The influences of a lambda prophage on the growth rate of Escherichia coli.

A lambda prophage increases the competitive ability of E. coli K12 in a glucose-limited chemostat. This phenomenon does not involve the lambdarex gene. Expression of the lambdarex gene conditionally decreases fitness in two ways: (1) it causes malT- but not malT+ lysogens to be at a selective disadvantage in competition with non-lysogens. (2) During adaptation to slow, glucose-limited chemostat growth from rapid, glucose-excess flask growth, expression of the lambdarex gene transiently decreases the growth rate when compared to the non-lysogen.

Bacteriophage lambda↗

[Differentiation of methicillin-resistant strains of Staphylococcus aureus by prophage specificity].

By inducing with mitomycin C the following phages were isolated from all the tested 32 methicillin resistant strains of S. aureus: the serogroup B phage was isolated from 2 strains, the serogroup B and F phages were isolated from 5 strains and the serogroup F phage was isolated from 25 strains. The phages were divided into 5 groups by the antiphage immunity. In group 1 of the phages 4 additional phages were specified. By the specificity of the prophages in the cultures all the strains were divided into 5 groups. Group 1 of the cultures was divided into 5 subgroups (A, B, C, D and E).

Bacteriophage Typing↗

[The effect of riboxine on prophage induction and the survival of bacterial cultures exposed to gamma irradiation].

In experiments with lysogenic cultures Pseudomonas aeruginosa and Escherichia coli K-12 proposed as a test for biological indication of relatively low doses of gamma-radiation, it was shown that when estimated by the criterion of prophage induction riboxine had a radioprotective action at nonlethal radiation doses. At the same time, when estimated by the criterion of the survival rate, riboxine produced the radioprotective effect at rather high radiation doses (survival rate of less than 10 per cent). It is suggested that riboxine is involved in the cell repair processes.

Bacteria↗

[Induction of prophage lambda during the irradiation of E. coli cells exposed to radiations of various LET].

Induction of lambda prophage in lysogenic E. coli cells exposed to ionizing radiation of different LET was studied as a function of dose I(D). Activities of pleiotropic RecA protein were shown to contribute to the shape of the I(D) curve. The experimental data were fitted by the function I(D) = alpha D(1-exp(-D0-1.D]exp(-beta D). Inducibility alpha increased with increasing LET which was related to the increased incidence of DNA lesions being a SOS-system call.

Bacteriophage lambda↗

[Transposition of the kanamycin resistance determinant (Tn601) from plasmid 5T to the genome of bacteriophage lambda and the expression of this gene after prophage induction].

Tn601, determinging kanamycin resistance of Escherichia coli, has been transposed into the bacteriophage lambda genome from R6 plasmid. After curing lambda gtc1857 (Tn601) lysogenes on the kanamycin containing medium, the clones with stable and unstable integrations of the Tn6-1 into the chromosome were obtained. After the lysogenization of these clones with the phage lambda att80c1857S7, the phages lambda att80c1857S7 (Tn601) were obtained. These phages contained the Tn601 from the sites of stable or unstable integrations. The frequency of the Tn601 transposition from the sites of unstable integration was 10(-7), that was two order of magnitude higher than the frequency of the Tn601 transpostion from the site of stable integration. Temperature induction of the lambda att80c1857 (Tn601) prophage resulted in 10--15 times increase of the yeild of aminoglycoside-3'-phosphotransferase I, the enzyme coded by the aphA gene of the Tn601.

Bacteriophage lambda↗

On the combined action of drugs on the lambda prophage induction in Escherichia coli K12 cells.

The capability of methotrexate, jododeoxyuridine and 5-fluorouracil to induce lambda prophage was compared when given alone or in combination. All these drugs were found to cause inducing conditions in Escherichia coli K12(lambda) cells. Combined action of jododeoxyuridine and methotrexate resulted in a pronounced increase in the number of free phages compared with that resulting the treatment either with methotrexate of jododeoxyuridine alone. Treatment with 5-fluorouracil caused inactivation of plaque forming ability in cells induced with methotrexate.

Coliphages↗

Electron microscopic study of phages and aberrant structures produced by induction of prophage kappa in Vibrio cholerae el tor cells.

Prophage kappa in V. cholerae el tor strain SLH22(J) could be induced spontaneously or by treatment with nitrofurantoin, though the efficiency of induction was very low (not more than 0.8%). V. cholerae el tor cells were found to release many different aberrant structures of the temperate phage, kappa. These aberrant structures were characterized by density gradient centrifugation and electron microscopy.

Bacteriophages↗

Population dynamics in the co-culture of Shigella flexneri 1b original strain and its antigenic 3b mutant carrying a prophage.

The antigenic mutant Shigella flexneri 3b showed selective prevalence when subcultured with the original strain 1b. Mathematical analysis of such co-cultures showed that the dynamics of bacterial growth may be described by equations of the Lotka-Volterra type. The analysis of serial cultivations suggests that parameters of the equations may be realizations of random variables characterizing strains and media. The mutant carries a prophage lethal for the original strain. Distinctive features of the growth of these strains alone and in co-culture may be successfully explained by the differences in growth parameters, without phage inference. The concurrence model connected only with the existence of the phage is not sufficient. A complete description of the dynamics of the co-culture is obtained by the connection of the assumptions of two models: strains in competition and growth of strains with phage, given the random character of the parameters.

Antigens, Bacterial↗

[Mucoid clones of Pseudomonas aeruginosa PAO1, surviving after induction of prophage transposons].

The origin and properties of mucoid clones were studied. The clones were selected with high frequency after thermo-induction of Pseudomonas aeruginosa lysogenic for phage transposons (PT). The production of alginate does not promote the survival of lysogenic bacteria at 42 degrees C. Mucoid clones were shown to appear before thermo-induction; the frequency of their formation does not depend on the specificity of the mutator effect intrinsic to different PT. Phenotypic differences typical of mucoid clones can be mediated by different mutations promoting clone survival at 42 degrees C and by simultaneously arising additional mutations. The SL21 mucoid clone selected among clones of P. aeruginosa PAO1 resistant to PT of B3 possesses an additional trait of phage resistance at 42 degrees C. The presence of D3112 cts 15 prophage has no significant effect on the frequency of SL21 reversion to nonmucoidness. This means that the mutator effect of PT has made a slight contribution to this process. The appearance of mutations promoting the survival of the thermoinducible lysogen SL21 (D3112 cts 15) does not affect the frequency of the loss of mucoidness. Nonmucoid derivatives of SL21 were shown to differ in phage resistance at 42 degrees C and in the extent of the residual mucoidness manifested under specific conditions. Consequently, nonmucoid clones appear as a result of pseudo-reversions. Because some of these pseudo-revertants cannot again be converted to the mucoid form, it is concluded that they carry mutations in genes whose functions are obligatory for the production of alginate.

Alginates↗