The Effect of 2,4-Dinitrophenol and Phage T2 on Escherichia Coli B

Heagy, F. C.

doi:10.1128/jb.59.3.367-373.1950

Cited by 25 publications

(11 citation statements)

References 6 publications

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“…Previously characterized LIN in E. coli phages is sensitive to changes in membrane proton motive force (PMF) and can be disrupted by the addition of energy poisons. One such poison, the ionophore 2,4-dinitrophenol (DNP), collapses the PMF and subsequently disrupts T2 and T4 LIN, resulting in rapid lysis of infected E. coli (28,29). To test if ICP1 LIN is similarly linked to PMF, we exposed PLE (-) V. cholerae infected at high MOI to DNP (Fig.…”

Section: Icp1 Causes Lysis Inhibition (Lin)mentioning

confidence: 99%

DominantVibrio choleraephage exhibits lysis inhibition sensitive to disruption by a defensive phage satellite

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2019

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AbstractBacteriophages and their bacterial hosts are locked in a dynamic evolutionary arms race. Phage satellites, selfish genomic islands which exploit both host bacterium and target phage, further complicate the evolutionary fray. One such tripartite system involves the etiological agent of the diarrheal disease cholera – Vibrio cholerae, the predominant phage isolated from cholera patients – ICP1, and a phage satellite – PLE. When ICP1 infects V. cholerae harboring the integrated PLE genome, PLE accelerates host lysis, spreading the PLE while completely blocking phage production protecting V. cholerae at the population level. Here we identify a single PLE gene, lidI, sufficient to mediate accelerated lysis during ICP1 infection and demonstrate that LidI functions through disrupting lysis inhibition – an understudied outcome of phage infection when phages vastly outnumber their hosts. This work identifies ICP1-encoded holin and antiholin genes teaA and arrA respectively, that mediate this first example of lysis inhibition outside the T-even coliphages. Through lysis inhibition disruption, LidI is sufficient to limit the number of progeny phage produced from an infection. Consequently, this disruption bottlenecks ICP1 evolution as probed by recombination and CRISPR-Cas targeting assays. These studies link novel characterization of the classic phenomenon of lysis inhibition with a conserved protein in a dominant phage satellite, highlighting the importance of lysis timing during infection and parasitization, as well as providing insight into the populations, relationships, and evolution of bacteria, phages, and phage satellites in nature.ImportanceWith increasing awareness of microbiota impacting human health comes intensified examination of, not only bacteria and the bacteriophages that prey upon them, but also the mobile genetic elements (MGEs) that mediate interactions between them. Research is unveiling evolutionary strategies dependent on sensing the milieu: quorum sensing impacts phage infection, phage teamwork overcomes bacterial defenses, and abortive infections sacrifice single cells protecting populations. Yet, the first discovered environmental sensing by phages, known as lysis inhibition (LIN), has only been studied in the limited context of T-even coliphages. Here we characterize LIN in the etiological agent of the diarrheal disease cholera, Vibrio cholerae, infected by a phage ubiquitous in clinical samples. Further, we show that a specific MGE, the phage satellite PLE, collapses LIN with a conserved protein during its anti-phage program. The insights gleaned from this work add to our expanding understanding of microbial fitness in natural contexts beyond the canonical bacterial genome and into the realm of antagonistic evolution driven by phages and satellites.

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Section: Icp1 Causes Lysis Inhibition (Lin)mentioning

confidence: 99%

DominantVibrio choleraephage exhibits lysis inhibition sensitive to disruption by a defensive phage satellite

Hays

Seed

2019

Preprint

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“…In normal invasion, this lyric reaction is soon halted (presumably after the DNA injection) and the leakage of cell constituents fails. These processes which normally check this reaction might themselves be inhibited by the attachment of an overwhelming dose of virus particles to the same cell, or by disturbance of the cell's metabolism by poisons (18,19), or nutrient deprivation (20). Heavily x-rayed phage may have suffered in its ability to carry out both the lyric and the subsequent lysis-inhibiting function, but if the latter were more readily damaged by irradiation, Watson's observations would be accounted for.…”

Section: Leakage Reactionmentioning

confidence: 99%

Mechanism of Cell Wall Penetration by Viruses

Puck

Lee

1954

Journal of Experimental Medicine

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The current status of delineation of the first phases of the invasive cycle of T2 bacteriophage is as follows: (a) Cell attachment is effected by binding of viral amino groups to complementarily spaced carboxyl groups on the cell surface (1, 2). (b) As a result of the establishment of these electrostatic bonds, the virus splits into its protein and DNA components (3, 2). (c) Only the DNA moiety penetrates into the cell, all or most of the protein remaining at the cell surface and apparently playing no further role in the subsequent reproductive steps (4).The mechanism of the reaction by which the DlffA is injected into the cell is still obscure. Mere splitting of the virus attached to the cell surface is not sufficient to cause DN'A penetration. Thus, when cell enzymes are inactivated by previous heating to 70°C., virus attachment and splitting still occur, but most of the DNA is liberated in free form into the medium instead of penetrating into the cell (5). Hence, the injection reaction appears to require action of cellular enzymes. Moreover, by manipulation of the salt concentration and the temperature, cell-attached virus can be induced to eject its DNA into the medium. Such virus does not contribute to cell killing (5). Since the cell-killing function has been shown to reside in the protein moiety alone (6), the conclusion seems warranted that DNA injection requires a triggering of cellular surface enzymes by the protein component of the virus, and that successful triggering is sufficient to cause death of the cell, even if the DNA does not afterwards enter. Finally, penetration is complete within 3 to 5 minutes at 37°C. because new specific synthesis of virus DNA inside the cell begins within this period (7). Hence, the penetration mechanism appears to be initiated by the protein component of the virus; its first steps are purely electrostatic, but it thereafter requires participation of enzymes in the cell surface; it leads to * This work is part of a research program carried out under Contract 1%. AT-(11-1)-269 with the Division of Biology and Medicine of the Atomic Energy Commission.Contribution No. 23.

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“…Phage was not synthesized and the infecting phage was lost. Heagy (1950) found that simultaneous infection with phage T2r+ and inhibition by 2,4-dinitrophenol under specified conditions is followed by immediate lysis of E. coli, strain B; the infecting phage is lost and no new phage is produced. In the cells of higher organisms at least, aerobic oxidation is mediated largely by enzyme systems in mitochondria; cyanide inhibits the cytochrome system and dinitrophenol inhibits oxidative phosphorylation.…”

Section: Discussionmentioning

confidence: 99%

Light and Electron Microscopic Studies of Escherichia Coli-Coliphage Interactions Iii

et al. 1953

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The first two articles of this series (Beutner et al., 1953; Mudd et al., 1953) contain descriptions of the light microscopic cytology and electron microscopy of T2 bacteriophage infection of Escherichia coli, strain B, with particular emphasis on the changes occurring in the chromatinic material, and with the intracellular appearance of possible phage precursors and apparently mature phage progeny. In this paper we show that the cytoplasmic organelles, the mitochondria, in contrast with the nuclei, maintain their integrity throughout the latent period of infection. MATERIALS AND METHODS Escherichia coli, strain B, and phages T2r+ and T2r were used throughout. The phage suspensions, media, basic techniques, and microscopic equipment have been described elsewhere (Beutner et al., 1953; Mudd et al., 1953). In addition, experiments were performed utilizing 2,3,5-triphenyltetrazolium chlorides as an indicator of reduction (Brodie and Gots, 1951, 1952; Kun, 1951; Shelton and Schneider, 1952). The reduction of 2,3,5-triphenyltetrazolium chloride was followed by mixing 3 ml aliquots of deepgrown (unaerated) log phase broth4 cultures of E. coli and 1 ml of a distilled water solution of I This work has been aided by a grant from the United States Atomic Energy Commission, AEC Contract no. AT(30-1)-1342. Aid was also received from the Theresa F. Felsen Memorial Fund.

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The Effect of 2,4-Dinitrophenol and Phage T2 on Escherichia Coli B

Cited by 25 publications

References 6 publications

DominantVibrio choleraephage exhibits lysis inhibition sensitive to disruption by a defensive phage satellite

DominantVibrio choleraephage exhibits lysis inhibition sensitive to disruption by a defensive phage satellite

Mechanism of Cell Wall Penetration by Viruses

Light and Electron Microscopic Studies of Escherichia Coli-Coliphage Interactions Iii

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