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PubMed1988

Bacteriophages of Bordetella sp.: features of lysogeny and conversion.

It has been the purpose of this paper to study molecular-biological features of the Bordetella bacteriophage interaction with the host cell during lysogeny and conversion as well as to determine the degree of homology between genomes of homologous and heterologous bacteriophages. Genomes of bacteriophages from B. pertussis 134, 41405 and B. bronchiseptica 214 were studied. Heteroduplex and restriction analyses revealed a heterogeneity of bacteriophage populations, and their DNAs were found to differ in size and position of inserts. As shown by blot hybridization, the bacteriophage genome is not inserted into the chromosome of the lysogenic cell but apparently exists as an autonomous plasmid replicon. It has been established that during conversion only a part of the phage genome is inserted into the chromosome of the recipient cell.

Autoradiography↗
PubMed2005

The genome sequence of Clostridium botulinum type C neurotoxin-converting phage and the molecular mechanisms of unstable lysogeny.

Botulinum neurotoxins (BoNTXs) produced by Clostridium botulinum are among the most poisonous substances known. Of the seven types of BoNTXs, genes for type C1 and D toxins (BoNTX/C1 and D) are carried by bacteriophages. The gene for exoenzyme C3 also resides on these phages. Here, we present the complete genome sequence of c-st, a representative of BoNTX/C1-converting phages. The genome is a linear double-stranded DNA of 185,682 bp with 404-bp terminal direct repeats, the largest known temperate phage genome. We identified 198 potential protein-coding regions, including the genes for production of BoNTX/C1 and exoenzyme C3. Very exceptionally, as a viable bacteriophage, a number of insertion sequences were found on the c-st genome. By analyzing the molecular structure of the c-st genome in lysogens, we also found that it exists as a circular plasmid prophage. These features account for the unstable lysogeny of BoNTX phages, which has historically been called "pseudolysogeny." The PCR scanning analysis of other BoNTX/C1 and D phages based on the c-st sequence further revealed that BoNTX phages comprise a divergent phage family, probably generated by exchanging genomic segments among BoNTX phages and their relatives.

Bacteriophages↗
PubMed1996

Purification and self-association equilibria of the lysis-lysogeny switch proteins of coliphage 186.

The CI repressor protein, responsible for maintenance of the lysogenic state, and the Apl protein, required for efficient prophage induction, are the two control proteins of the lysis-lysogeny transcriptional switch of coliphage 186. These proteins have been overexpressed, purified, and their self-association behavior examined by sedimentation equilibrium. Phage 186 CI dimers self-associate in solution through tetramers to octamers in a concerted process. The Apl protein of 186 is an unusual example of a helix-turn-helix protein which is monomeric in solution.

Base Sequence↗
PubMed2000

Establishment of lysogeny in bacteriophage 186. DNA binding and transcriptional activation by the CII protein.

The CII protein of bacteriophage 186 is a transcriptional activator of the helix-turn helix family required for establishment of the lysogenic state. DNA binding by 186 CII is unusual in that the invertedly repeated half sites are separated by 20 base pairs, or two turns of the DNA helix, rather than the one turn usually associated with this class of proteins. Here, we investigate quantitatively the DNA binding properties of CII and its interaction with RNA polymerase at the establishment promoter, p(E). The stoichiometry of CII binding was determined by sedimentation equilibrium experiments using a fluorescein-labeled oligonucleotide and purified CII. These experiments indicate that the CII species bound to DNA is a dimer, with additional weak binding of a tetrameric species at high concentrations. Examination of the thermodynamic linkages between CII self-association and DNA binding shows that CII binds to the DNA as a preformed dimer (binding free energy, 9.9 kcal/mol at 4 degrees C) rather than by association of monomers on the DNA. CII binding induces in the DNA a bend of 41 (+/- 5) degrees. The spacing between the binding half sites was shown to be important for CII binding, insertion or removal of just 1 base pair significantly reducing the affinity for CII. Removal of 5 or 10 base pairs between binding half sites eliminated binding, as did insertion of an additional 10 base pairs. CII binding at p(E) was improved marginally by the presence of RNA polymerase (DeltaDeltaG = -0.5 (+/- 0.3) kcal/mol). In contrast, the binding of RNA polymerase at p(E) was undetectable in the absence of CII but was improved markedly by the presence of CII. Thus, CII appears to recruit RNA polymerase to the promoter. The nature of the base pair changes in mutant phage, selected by their inability to establish lysogeny, are consistent with this mechanism of CII action.

Base Sequence↗
PubMed1975

The nature and incidence of lysogeny in Mycobacterium fortuitum.

Ten of 28 strain of Mycobacterium fortuitum (ranae) were found to be associated with bacteriophage; three were pseudolysogenic, one liberated a phage that lysed a sensitive indicator strain, two liberated morphologically complete phages that did not lyse any of the strains used in this study and four liberated morphologically defective phages. The lysogenic and defectively lysogenic strains showed anomalies in cultural, biochemical and antigenic properties and in susceptibility to superinfecting phages. In view of the high frequency of lysogeny found in M. fortuitum, the role of bacteriophage in the variation of properties, including pathogenicity, of mycobacteria of greater clinical importance merits further consideration.

Bacteriolysis↗
PubMed2003

Lysogeny and bacteriophage host range within the Burkholderia cepacia complex.

The Burkholderia cepacia complex comprises a group of nine closely related species that have emerged as life-threatening pulmonary pathogens in immunocompromised patients, particularly individuals with cystic fibrosis or chronic granulomatous disease. Attempts to explain the genomic plasticity, adaptability and virulence of the complex have paid little attention to bacteriophages, particularly the potential contribution of lysogenic conversion and transduction. In this study, lysogeny was observed in 10 of 20 representative strains of the B. cepacia complex. Three temperate phages and five lytic phages isolated from soils, river sediments or the plant rhizosphere were chosen for further study. Six phages exhibited T-even morphology and two were lambda-like. The host range of individual phages, when tested against 66 strains of the B. cepacia complex and a representative panel of other pseudomonads, was not species-specific within the B. cepacia complex and, in some phages, included Burkholderia gladioli and Pseudomonas aeruginosa. These new data indicate a potential role for phages of the B. cepacia complex in the evolution of these soil bacteria as pathogens of plants, humans and animals, and as novel therapeutic agents.

Bacteriophages↗
PubMed2001

Octamerization of lambda CI repressor is needed for effective repression of P(RM) and efficient switching from lysogeny.

The CI repressor of bacteriophage lambda is a model for the role of cooperativity in the efficient functioning of genetic switches. Pairs of CI dimers interact to cooperatively occupy adjacent operator sites at O(R) and at O(L). These CI tetramers repress the lytic promoters and activate transcription of the cI gene from P(RM). CI is also able to octamerize, forming a large DNA loop between O(R) and O(L), but the physiological role of this is unclear. Another puzzle is that, although a dimer of CI is able to repress P(RM) by binding to the third operator at O(R), O(R)3, this binding seems too weak to affect CI production in the lysogenic state. Here we show that repression of P(RM) at lysogenic CI concentrations is absolutely dependent on O(L), in this case 3.8 kb away. A mutant defective in this CI negative autoregulation forms a lysogen with elevated CI levels that cannot efficiently switch from lysogeny to lytic development. Our results invalidate previous evidence that Cro binding to O(R)3 is important in prophage induction. We propose the octameric CI:O(R)-O(L) complex increases the affinity of CI for O(R)3 by allowing a CI tetramer to link O(R)3 and the third operator at O(L), O(L)3.

Bacteriophage lambda↗
PubMed1990

Temperate bacteriophages and lysogeny in lactic acid bacteria.

Lysogeny is widespread in the lactic acid bacteria. The majority of lysogens can be induced by UV irradiation or treatment with mitomycin C, but indicator strains which allow lytic growth of the induced phage are often not easy to identify. A few temperate phages have been shown to transduce chromosomal and/or plasmid markers. Information about the molecular biology of the temperate phages from lactic acid bacteria is sparse and needs significant supplementation in order that these potentially valuable phages might be utilized more efficiently as tools for improving existing starter strains in dairy fermentations.

Amino Acid Sequence↗
PubMed1964

SEQUENTIAL GENE ACTION IN THE ESTABLISHMENT OF LYSOGENY.

A study of temperature sensitive clear-plaque-forming mutants of bacteriophage P22 demonstrates that the c(1) and c(2) loci must function in a temporal sequence in the establishment of lysogeny in Salmonella typhimurium. The c(1) locus acts for only a 4-minute interval between the 7th to 11th minute of the infection. The c(2) locus begins to function some minutes later, and its continued activity is necessary for perpetuation of the lysogenic state.

DNA↗
PubMed1992

Lysogeny and bacteriocinogeny in Xenorhabdus nematophilus and other Xenorhabdus spp.

Induction by mitomycin or high-temperature treatment resulted in the production of bacteriocins and phages in both phases of Xenorhabdus nematophilus A24, indicating lysogeny. Phage DNA purified from X. nematophilus A24 hybridized to several fragments of DraI-digested A24 chromosomal DNA, confirming that the phage genome was incorporated into the bacterial chromosome. Bacteriocins and phages were detected in cultures of most other Xenorhabdus spp. after mitomycin or high-temperature treatment. Xenorhabdus luminescens K80 was not lysed by these treatments, and no phages were seen associated with this strain. However, bacteriocins were detected in limited quantities in all Xenorhabdus cultures, including X. luminescens K80, without any induction. X. nematophilus A24 bacteriocins were antagonistic for other Xenorhabdus species but not for A24 or other strains of X. nematophilus.

Bacteriocins↗
PubMed2002

Seasonal variation in lysogeny as depicted by prophage induction in Tampa Bay, Florida.

A seasonal study of the distribution of lysogenic bacteria in Tampa Bay, Florida, was conducted over a 13-month period. Biweekly water samples were collected and either were left unaltered or had the viral population reduced by filtration (pore size, 0.2 micro m) and resuspension in filtered (pore size, 0.2 micro m) water. Virus-reduced and unaltered samples were then treated by adding mitomycin C (0.5 micro g ml(-1)) to induce prophage or were left untreated. In order to test the hypothesis that prophage induction was phosphate limited, additional induction experiments were performed in the presence and absence of phosphate. Induction was assessed as an increase in viral direct counts, relative to those obtained in controls, as detected by epifluorescence microscopy. Induction of prophage was observed in 5 of 25 (20%) unaltered samples which were obtained during or after the month of February, paralleling the results from a previous seasonal study. Induction of prophage was observed in 9 of 25 (36%) of the virus-reduced samples, primarily those obtained in the winter months, which was not observed in a prior seasonal study (P. K. Cochran and J. H. Paul, Appl. Environ. Microbiol. 64:2308-2312, 1998). Induction was noted in the months of lowest bacterial and primary production, suggesting that lysogeny was favored under conditions of poor host growth. Phosphate addition enabled prophage induction in two of nine (22%) experiments. These results indicate that prophage induction may occasionally be phosphate limited or respond to increases in phosphate concentration, suggesting that phosphate concentration may modulate the lysogenic response of natural populations.

Bacteria↗
PubMed1984

Effect of bacteriophage P1 lysogeny on lipopolysaccharide composition and the lambda receptor of Escherichia coli.

The outer membrane of Escherichia coli was altered as a consequence of lysogeny by bacteriophages P1 and P1 cmts. The predominant change was a reduction in the size of lipopolysaccharide to a heptose-deficient form. P1 cmts lysogens were still sensitive to several bacteriophages but were resistant to lambda vir. Neither whole cells nor solubilized outer membranes from P1 cmts lysogens were able to inactivate lambda vir, and 32P-labeled lambda vir was unable to adsorb to P1 cmts lysogens. P1 cmts lysogens were also affected in maltose transport. The level of periplasmic maltose-binding protein was reduced somewhat, but there was no significant reduction in the level of the outer membrane lambda receptor (LamB). These membrane abnormalities were all corrected in strains cured of P1 cmts. It is suggested that P1 cmts affects lipopolysaccharide biosynthesis by a phage conversion mechanism and consequently the function of the lambda receptor.

Bacterial Outer Membrane Proteins↗
PubMed1993

Involvement of Escherichia coli FIS protein in maintenance of bacteriophage mu lysogeny by the repressor: control of early transcription and inhibition of transposition.

The Escherichia coli FIS (factor for inversion stimulation) protein has been implicated in assisting bacteriophage Mu repressor, c, in maintaining the lysogenic state under certain conditions. In a fis strain, a temperature-inducible Mucts62 prophage is induced at lower temperatures than in a wild-type host (M. Bétermier, V. Lefrère, C. Koch, R. Alazard, and M. Chandler, Mol. Microbiol. 3:459-468, 1989). Increasing the prophage copy number rendered Mucts62 less sensitive to this effect of the fis mutation, which thus seems to depend critically on the level of repressor activity. The present study also provides evidence that FIS affects the control of Mu gene expression and transposition. As judged by the use of lac transcriptional fusions, repression of early transcription was reduced three- to fourfold in a fis background, and this could be compensated by an increase in cts62 gene copy number. c was also shown to inhibit Mu transposition two- to fourfold less strongly in a fis host. These modulatory effects, however, could not be correlated to sequence-specific binding of FIS to the Mu genome, in particular to the strong site previously identified on the left end. We therefore speculate that a more general function of FIS is responsible for the observed modulation of Mu lysogeny.

Bacteriophage mu↗
PubMed1961

Lysogeny in staphylococci.

Blair, John E. (Hospital for Joint Diseases, New York, N. Y.) and Miriam Carr. Lysogeny in staphylococci. J. Bacteriol. 82:984-993. 1961.-Changes in the phage typing patterns of strains of staphylococci of the 80/81-52/52A/80/81 complex and of phage group III were produced by lysogenization with temperate phages derived from selected strains of Staphylococcus aureus. The phages used were of the serological groups A, B, and F. Certain changes of phage pattern were related to serologically specific prophage immunity; others were nonspecific, or resulted from the conversion of a strain from partial resistance to full sensitivity to certain typing phages. In addition to an alteration of their phage typing pattern, the lysogenization of certain strains by appropriate phages effected a reversal of their susceptibility to penicillin. The capacity to produce toxin was conferred upon certain nontoxigenic strains by lysogenization with a phage derived from a toxigenic strain.

Bacteriophage Typing↗
PubMed1963

DEMONSTRATION OF LYSOGENY IN STABLE L FORMS OF PROTEUS MIRABILIS.

Taubeneck, Udo (Institut für Mikrobiologie und Experimentelle Therapie, Jena, Germany). Demonstration of lysogeny in stable L forms of Proteus mirabilis. J. Bacteriol. 86:1265-1269. 1963.-Stable L forms of eight different lysogenic Proteus mirabilis strains have proved to be lysogenic. In an additional strain, phage production was induced upon transfer into the stable L form. This demonstrates that the stable L forms, like their parent bacillary types, are capable of forming mature phage. Their phage resistance is not due to an inability to form mature phage but to a loss of phage receptors. In some instances, the transition into the stable L form is associated with altered phage production, usually with increased phage production, in comparison with the parental bacillary type.

Bacillus↗
PubMed1965

EVIDENCE OF LYSOGENY IN DERIVATIVES OF ESCHERICHIA COLI.

Frampton, E. W. (The University of Texas M. D. Anderson Hospital and Tumor Institute, Houston), and B. R. Brinkley. Evidence of lysogeny in derivatives of Escherichia coli. J. Bacteriol. 90:446-452. 1965.-Exposure of cultures of several thymine-requiring derivatives of Escherichia coli strain 15 to radiation [X rays or ultraviolet (UV)] resulted in an increased synthesis of ribonucleic acid (RNA) as measured by the incorporation of radioactive uridine. In addition to radiation, mitomycin C (5 or 25 mug/ml) or growth in the absence of thymine also stimulated RNA synthesis in cultures in which the bulk of RNA synthesis was terminated by a shift-down in growth conditions. RNA purified from X-irradiated bacteria was resolved on columns of methylated serum albumin and yielded an elution profile which appeared similar to RNA synthesized by phage-infected bacteria. Electron microscopic examination of cultures induced to lyse by exposure to UV radiation, or by growth in the presence of 5-bromouracil before exposure to radiation (X rays or UV), showed the presence of mostly incomplete phage particles.

Bacteriolysis↗
PubMed1968

Effect of lysogeny on serum sensitivity.

When Escherichia coli K-12 was infected with lambda phage and mutants of lambda characterized by the production of temperature-sensitive repressors, the lysogenic bacteria were significantly more resistant to normal serum than the uninfected organisms. Infection of E. coli K-12 with a lambdoid phage, phi80, whose prophage attachment site is different from that of lambda, did not result in a detectable change in serum resistance. Similarly, infection with certain Pseudomonas and Shigella phages caused no detectable differences in serum resistance. Finally, the well-known conversion of the Salmonella anatum serotype to S. newington by E(15) phage indicated that, despite the relatively greater roughness of S. anatum, S. newington was more sensitive to normal serum than S. anatum. Thus, the effects of lysogeny on the sensitivity of bacteria to the bactericidal action of serum mediated by the complement system may be quite variable.

Bacteriophages↗
PubMed1975

Chloramphenicol stimulation of lysogeny by lambda regulatory mutants.

Inhibition of protein synthesis in Escherichia coli by amino acid starvation or chloramphenicol addition increases the frequency of lysogeny by lambda phage two- to fourfold. Lambda cIII mutants, which normally lysogenize at very low frequencies, lysogenize at very high frequencies in the presence of chloramphenicol.

Bacterial Proteins↗