Bacteriophage T5 Chromosome Fractionation: Genetic Specificity of a DNA Fragment

Lanni, Yvonne Théry; Lanni, Frank; Tevethia, Mary J.

doi:10.1126/science.152.3719.208

Cited by 23 publications

(16 citation statements)

References 10 publications

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“…E. coli F is a fast-adsorbing host strain for phage T5 (38). E. coli HR34 (39) is the isogenic Sup' strain and allows the growth of the T5 A2-(amlO2) amber mutant (40). E. coli K-12 was from our collection.…”

Section: Methodsmentioning

confidence: 99%

“…Indeed, Lanni showed (37) and it was verified again recently in our laboratory (20, 21) that superinfecting Alor A2am mutants can efficiently use proteins Al and A2 previously synthesized by wild-type FST DNA to inject their own complete DNA without arrest at the FST stage. Lanni also showed that part of the entering T5 A2-DNA can recombine with wildtype FST DNA to produce T5+ phage (40). These two methods of production of phage in sup bacteria, which were detected by plating on sup' and sup-lawns, were used as tests of total injection of T5 DNA.…”

Section: Concentrated Bacteria (5 X 109 Cells Per Ml)mentioning

confidence: 99%

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Host cell metabolic energy is not required for injection of bacteriophage T5 DNA

Maltouf¹,

Labedan²

1983

J Bacteriol

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The addition of various metabolic inhibitors (uncouplers, cyanide, arsenate, ionophores) separately or together (for example, arsenate and an uncoupler) or even harsher methods of energy depletion did not prevent bacteriophage T5 from injecting its first-step-transfer DNA (a DNA segment 3 ,um long) into the cytoplasm of host cells. The same indifference to metabolic energy was observed if first-step-transfer DNA was decapsidated and uncoiled before injection, thus precluding any energetic help from the phage capsid or from some tension stored in DNA tightly packed in the head. Penetration of the second-step-transfer DNA across the cytoplasmic membrane was studied by determining injection of superinfecting T5 A2-amber phages into Sup-bacteria containing proteins Al and A2 previously encoded by the first-step-transfer DNA of a primary wild-type phage. The addition of various metabolic inhibitors after synthesis of proteins Al and A2 but before superinfection did not prevent this penetration of second-steptransfer DNA. Thus, we conclude that traversal of the cytoplasmic membrane by the entire T5 DNA (a molecule 34 ,m long) must occur by diffusion through protein channels.

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Section: Methodsmentioning

confidence: 99%

Section: Concentrated Bacteria (5 X 109 Cells Per Ml)mentioning

confidence: 99%

Host cell metabolic energy is not required for injection of bacteriophage T5 DNA

Maltouf¹,

Labedan²

1983

J Bacteriol

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“…The T5 ami5 mutant is unable to make the transfer protein necessary for the injection of post-FST DNA and to degrade the bacterial DNA (7). It was isolated by Lanni et al (8) and normally grows on E. coli HR34 (su+) but not in the F (suj) strain (7). This mutant was used in the same type of experiments we have already described to verify that the transfer of naked post-FST DNA requires the presence of the transfer protein.…”

Section: Resultsmentioning

confidence: 99%

Penetration into Host Cells of Naked, Partially Injected (Post-FST) DNA of Bacteriophage T5

Labedan

Legault-Démare

1973

J Virol

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The experiments presented here suggest that the naked post-FST DNA, attached to the bacteria after a centrifugation at 6,000 × g of Escherichia coli -T5 complexes arrested at the FST stage, can be injected into the host cell if protein synthesis is allowed. This results from the observation that the production of infectious centers by this naked DNA is sensitive to DNase or moderate shearing treatments. The process of injection seems to be the same as that for DNA normally rolled in an adsorbed capsid.

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“…This slowness is perhaps attributable to complexity of this phenomenon. In the case of T5 phage (B-group) active against Escherichia coli, for full injection of DNA there must first be injection of first-step-transfer DNA, then expression of early genes, coded on first-step-transfer DNA, and only then injection of the remainder of the DNA (Lanni et al 1964;McCorquodale & Lanni, 1964;Lanni, 1965Lanni, , 1969Lanni & Lanni, 1966;Lanni et al I966;McCorquodale & Buchanan, 1968). For the present, however, it is not certain whether this mechanism is true also for PL-I, although even in the case of T5 a lot of ATP might be required along the way.…”

Section: Discussionmentioning

confidence: 99%

Adenosine Triphosphate Content in Lactobacillus casei and the Blender-resistant Phage-Cell Complex-forming Ability of Cells on Infection with PL-1 Phage

Watanabe

Takesue

Ishibashi

1979

Journal of General Virology

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SUMMARYThe intracellular ATP content of Lactobacillus casei ATCC 27o92 grown in a glucose-containing medium was almost constant (2 to 3/zg/mg dry wt. cells) through the early to middle stage of logarithmic phase, but it was lowered to less than o. I #g/mg after cessation of growth owing to the exhaustion of available glucose. All the cells in the early stage of stationary phase were still viable and thus considered to be in a starved state. When such starved cells were infected with PL-~ phages in a tris-maleate buffer of pH 6.0, the process of forming blender-resistant phagecell complexes signifying the complete injection of phage genomes into the cells was much inhibited. There was a good correlation between the ATP content of cells and the extent of the formation of blender-resistant phage-cell complexes and the correlation coefficient between them was 0"89 + o'o9 at the 95 % confidence limit. On the other hand, the process of forming both the phage-adsorbed cells and the anti-phage serum-resistant phage-cell complexes were not affected by the ATP content of cells. Feeding of glucose to such starved cell cultures caused the ceils to restore both the ATP content and the ability to form blender-resistant phage-ceil complexes. Such restoration was also observed when the starved cells collected by centrifugation were incubated in a glucose-containing medium. The significance of the intracellular level of high energy compounds such as ATP for the mechanism of the injection of phage genomes into the cells is discussed.

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Bacteriophage T5 Chromosome Fractionation: Genetic Specificity of a DNA Fragment

Cited by 23 publications

References 10 publications

Host cell metabolic energy is not required for injection of bacteriophage T5 DNA

Host cell metabolic energy is not required for injection of bacteriophage T5 DNA

Penetration into Host Cells of Naked, Partially Injected (Post-FST) DNA of Bacteriophage T5

Adenosine Triphosphate Content in Lactobacillus casei and the Blender-resistant Phage-Cell Complex-forming Ability of Cells on Infection with PL-1 Phage

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