ANALYSIS OF A GENE CONTROLLING CELL DIVISION AND SENSITIVITY TO RADIATION IN
            <i>ESCHERICHIA COLI</i>

Adler, Howard I.; Hardigree, A.A.

doi:10.1128/jb.87.3.720-726.1964

Cited by 159 publications

(61 citation statements)

References 3 publications

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“…Therefore, JC355 was treated with NTG, and a mucoid mutant (ES370) was selected on minimal medium at 30 C as described above. This mutant was Plkc sensitive and also T7 sensitive, The mutation to mucoid (excess capsule formation) can occur at two distinct loci: capR (synonomous with Ion; see references 3,14,19) and capS (21). The capR mutants are also UV sensitive (3,9,14).…”

Section: Resultsmentioning

confidence: 94%

Mutation Preventing Capsular Polysaccharide Synthesis in Escherichia coli K-12 and Its Effect on Bacteriophage Resistance

Radke

Siegel

1971

J Bacteriol

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A mutant strain of Escherichia coli K-12 was found in which spontaneous mutation to phage T7 resistance occurred at a very low frequency. T7 resistance in the parental strain from which this mutant was derived resulted from a mutation to excess capsular polysaccharide synthesis. non-9, inhibited capsule formation when non-9 mutation was cotransducible with non-9 his Su-l.The mutation preventing T7 resistance, transduced into capsulated strains. The his, the gene order in this region being The T phages of Escherichia coli have in common the ability to adsorb to and replicate in E. coli strains B and K-12. Compared with many E. coli strains found in nature, strains B and K-12 are "rough," i.e., they do not have significant amounts of extracellular polysaccharide capsular material. The T phages adsorb to specific sites on the cell wall (30). Mutants of E. coli K-12 and other strains of E. coli are found that produce very large amounts of capsular material and are mucoid or "smooth" in appearance (16,19,24). Some, but not all of these mutants, are resistant to the T phages and other coliphages because the cell wall phage receptors are covered by the capsule polysaccharide (16, 24).We have found a mutant of E. coli K-12 that mutates at a very low frequency to resistance to phage T7. This work will show that this unusual phenotype is due to a block in capsule formation.MATERIALS AND METHODS Bacterial strains, phages, media, and growth conditions. The bacterial strains used are listed in Table 1. Wild-type T phages were obtained from laboratory stocks and grown and assayed on E. coli B by using procedures described by Adams (1). The methods for P 1 kc growth and transduction were previously described (27).The minimal medium was that of Davis and Mingioli (6) plus I ug of thiamine hydrochloride and 0.5 sg of ferrous sulfate per ml. When necessary, amino acids were used at a final concentration of 50 tig/ml. Nutrient broth contained 0.8% (w/v) nutrient broth powder (Difco), 0.1% yeast extract (Difco), and 0.5% sodium chloride. Solid media (nutrient and minimal agar) were made by adding 20 g of agar (Difco) per liter to the liquid medium. Antibiotic medium no. 3 (Difco) was occasionally used in place of nutrient broth. Bromothymol blue medium, used to score fermentation markers, contained 0.8% Difco nutrient broth powder, 1.0% sugar, 0.006% bromothymol blue, and 2% agar.Overnight, aerated cultures grown at 37 C were used as stationary-phase cultures. Liquid cultures and plates were incubated at 30 C when necessary to increase capsule formation; all other incubations were at 37 C. Genetic procedures. All matings were uninterrupted. Donor and recipient cultures were mixed and incubated for I to 2 hr before plating as described (27). In transductions, a multiplicity of infection of one was used. All recombinants were purified by single-colony isolation on the selective medium.Unselected auxotrophic and fermentation markers were scored by replica plating. The Su-l+ (suppressor containing) and Su-1alleles were scored by crossst...

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Section: Resultsmentioning

confidence: 94%

Mutation Preventing Capsular Polysaccharide Synthesis in Escherichia coli K-12 and Its Effect on Bacteriophage Resistance

Radke

Siegel

1971

J Bacteriol

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“…11. Pantoyl lactone is a division-promoting agent (2,10). Its effect on colony survival can give information about the effect of filament formation on colonyforming ability.…”

Section: Resultsmentioning

confidence: 99%

Effect of Nalidixic Acid and Hydroxyurea on Division Ability ofEscherichia coli fil⁺andlon⁻Strains

Kantor

Deering

1968

J Bacteriol

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Short periods of incubation in medium containing nalidixic acid or hydroxyurea, followed by a return to normal growth conditions, induced filament formation in Escherichia coli B (fil+) and AB1899NM (Ion-) but not in B/r (fil-) and AB1157 (lon+). These drugs reversibly stopped deoxyribonucleic acid (DNA) synthesis with little or no effect on ribonucleic acid (RNA) synthesis or mass increase. The initial imbalance caused by incubation in these drugs was the same for B and B/r as was macromolecular synthesis following a return to normal growth conditions. DNA degradation caused by nalidixic acid was measured and found to be the same for B and B/r. Hydroxyurea caused no DNA degradation in these two strains. Survival curves as determined under various conditions by colony formation suggested that the property of filament formation was responsible for the extrasensitivity of fil+ and lonstrains to either nalidixic acid or hydroxyurea. E. coli B was more sensitive to either drug than was B/r or B,-1. Pantoyl lactone or liquid holding treatment aided division and colony formation of nalidixic acid-treated B but had no effect on B/r. Likewise, the filament-former AB1899NM was more sensitive to nalidixic acid than was the non-filament-former ABI 157. The sensitivity of B/r and B.,1 to nalidixic acid was nearly the same except at longer times in nalidixic acid, when B8-1 appeared more resistant. Even though nalidixic acid, hydroxyurea, and ultraviolet light may produce quite different molecular alterations in E. coli, they all cause a metabolic imbalance resulting in a lowered ratio of DNA to RNA and protein. We propose that it is this imbalance per se rather than any specific primary chemical or photochemical alterations which leads to filament formation by some genetically susceptible bacterial strains such as lon-and fil+.Certain strains of Escherichia coli, referred to as MATERIALS AND METHODS Bacterial strains and growth conditions. The bacterial strains used in this investigation were E. coli B, 520 on July 10, 2020 by guest

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“…Hirota et al (16) have mapped many division mutants of Escherichia coli and has found no less than seven genetic loci which affect cell division. Others have isolated mutants of cell division (2,3,6,11,16,17,21,23,26,31,32), and in some cases the lesions have been mapped (1,18).…”

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Regulation of Cell Division inEscherichia coli:Characterization of Temperature-Sensitive Division Mutants

1970

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A temperature-sensitive division mutant of Escherichia coli was isolated by using differential filtration to select for filaments at 42 C and normal cells at 30 C. Cells shifted from 30 to 42 C stop dividing almost immediately, suggesting the temperature-sensitive element is required for cell division late in the cell cycle. Cells returned to 30 from 42 C divide abruptly, suggesting accumulation of division potential at 42 C. Inhibitors of protein, deoxyribonucleic acid, and ribonucleic acid synthesis do not block division during the recovery period at 30 C. Cycloserine does not stop cell division, vancomycin shows some effect on cell division, whereas penicillin completely stops cell division during this period. The addition of high concentrations of NaCl to filaments at 42 C results in a burst of cell division. The final cell number is equivalent to the control which is grown at 30 C if sufficient salt is added (11 g/liter, final concentration). After the original burst, cell division ceases at the nonpermissive temperature even at increased osmolality. Chloramphenicol, puromycin, vancomycin, and penicillin prevent division during the recovery in the presence of NaCl. Kinetic data indicate division potential decays to a reversible inactive intermediate which rapidly decays to an irreversible inactive form. Conversion of division potential to the inactive form is correlated with a 100to 1,000-fold derepression of the synthesis of division potential. The mutation appears to involve a stage in cross-wall synthesis which is required during the terminal stages of division.

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ANALYSIS OF A GENE CONTROLLING CELL DIVISION AND SENSITIVITY TO RADIATION IN ESCHERICHIA COLI

Cited by 159 publications

References 3 publications

Mutation Preventing Capsular Polysaccharide Synthesis in Escherichia coli K-12 and Its Effect on Bacteriophage Resistance

Mutation Preventing Capsular Polysaccharide Synthesis in Escherichia coli K-12 and Its Effect on Bacteriophage Resistance

Effect of Nalidixic Acid and Hydroxyurea on Division Ability ofEscherichia coli fil⁺andlon⁻Strains

Regulation of Cell Division inEscherichia coli:Characterization of Temperature-Sensitive Division Mutants

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ANALYSIS OF A GENE CONTROLLING CELL DIVISION AND SENSITIVITY TO RADIATION IN ESCHERICHIA COLI

Cited by 159 publications

References 3 publications

Mutation Preventing Capsular Polysaccharide Synthesis in Escherichia coli K-12 and Its Effect on Bacteriophage Resistance

Mutation Preventing Capsular Polysaccharide Synthesis in Escherichia coli K-12 and Its Effect on Bacteriophage Resistance

Effect of Nalidixic Acid and Hydroxyurea on Division Ability ofEscherichia coli fil+andlon−Strains

Regulation of Cell Division inEscherichia coli:Characterization of Temperature-Sensitive Division Mutants

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Effect of Nalidixic Acid and Hydroxyurea on Division Ability ofEscherichia coli fil⁺andlon⁻Strains