Resporulation of outgrowing Bacillus subtilis spores

Keynan, A.; Berns, A A; Dunn, G.; Young, Michael; Mandelstam, J.

doi:10.1128/jb.128.1.8-14.1976

Cited by 20 publications

(12 citation statements)

References 22 publications

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“…Sporulation is a multiphase process leading to the development of a spore from a vegetative cell. The stages involved (57,118,119) can be summarized as follows. Stage 0 is the vegetative cell, stage I the presporulation phase (in which DNA is present as an axial filament), and stage II is the septation phase in which asymmetric cell formation occurs.…”

Section: Sporulationmentioning

confidence: 99%

Bacterial spores and chemical sporicidal agents.

Russell¹

1990

Clinical Microbiology Reviews

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Section: Sporulationmentioning

confidence: 99%

Bacterial spores and chemical sporicidal agents.

Russell¹

1990

Clinical Microbiology Reviews

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“…Germination and outgrowth. Germination and outgrowth of approximately 2 x i0W spores per ml were carried out in Difco Penassay broth (supplemented with 20,ug of tryptophan per ml) or in similarly supplemented brain heart infusion broth (Difco), containing 5 g of yeast extract (Difco) per liter (13), and incubated at 370C on a rotatory shaker (250 rpm). When indicated, the spore suspension was heat activated for 7 min at 800C before incubation at 370C (zero time).…”

Section: Ur-mentioning

confidence: 99%

“…Septa. Septa were counted by phase-contrast microscopy after mixing 1 drop of the cell suspension with 1 drop of saturated CsCl (13).…”

Section: Ur-mentioning

confidence: 99%

“…Sequential release of DNA during cell outgrowth. Optimum conditions for synchrony of germination and outgrowth were found by using the very rich medium consisting of brain heart infusion with yeast extract previously used by Keynan et al (13). These conditions led to rapid germination, as revealed by phase darkening of 93% of the spores during the first 15 min and appearance of septa beginning at 87 min, with 90% of the cells showing one or more septa 23 min later (Fig.…”

Section: Ur-mentioning

confidence: 99%

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Bacillus subtilis 168 genetic transformation mediated by outgrowing spores: necessity for cell contact

Orrego¹,

Arnaud²,

Halvorsen³

1978

J Bacteriol

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Transforming activity released in sequential genetic order during the first synchronous cycle of DNA replication during outgrowth of spores of Bacillus subtilis 168 was investigated. A transformation assay was used consisting of outgrowing spores as DNA donors and multiply marked competent cells as recipients. DNA synthesis inhibitors known to stop DNA release were used during and subsequent to DNA transfer to recipient cells. The released DNA sedimented with the outgrowing cells after low-speed centrifugation, and it was discovered that markers released both early and late were resistant to up to 500 pg of deoxyribonuclease per ml under conditions in which the transforming capacity of purified DNA was eliminated by 5 ,ug of the nuclease per ml. Inaccessibility to deoxyribonuclease was increased and maintained during the transfornation event while detergents and proteolytic attack did not expose the released chromosome to nuclease action. The results indicate that tight physical contact between outgrowing spores and competent cells is required for transformation in this system.Borenstein and Ephrati-Elizur (2) discovered that outgrowing cells originating from a culture of dormant Bacillus subtilis 168 spores release DNA in a sequential genetic order and in synchrony with the first replication cycle of the genome. Release of genetic material was followed by simply mixing samples from an outgrowing culture with appropriately marked competent celLs. Markers closer to the origin of replication were transformed first, whereas markers away from the origin were ransformed later, as a function of DNA replication time. These observations were used for genetic mapping (2). The generality of these original observations is nevertheless in question, given that nonsequential release of certain markers firom particular B. subtilis strains has been observed (W. D. Crabb and U. Streips, Abstr. Annu. Meet. Am. Soc. Microbiol. 1976, H8, p. 97), suggesting different replication patterns between strains or a lag between replication and release which varies between chromosomal regions. A correlation between DNA synthesis and DNA release was indicated by the fact that inhibition of chromosome replication in the outgrowing culture impaired inhibition of transfonation ofcompetent cells (2). These observations, coupled with the sequential nature of the release, excluded the possibility that the transformation observed was due to lysis of the outgrowing cells, either in the liquid and/or during their residence on the agar plate after mixing with the comptent cells. ur-The physical nature of the DNA release during spore outgrowth has not been explored. This communication presents evidence that the released copy of the bacterial chromosome remains attached to the ceil in a form inaccessible to deoxyribonuclease (DNase) yet available for transformation of competent cells. The results obtained indicate that tight physical contact between outgrowing spores and competent cells is required for transformation in this system.(Parts of this report h...

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“…In recent years, evidence has accumulated that induction of sporulation in Bacillus sp. can occur only during a limited stage in the cell cycle and is linked to DNA replication (11,15). Evidence for a link between the DNA replication cycle and induction of differentiation has also been demonstrated in a variety of other differentiating cells (9; for reviews, see also 17).…”

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confidence: 99%

Independence of Bacillus subtilis spore outgrowth from DNA synthesis

Ginsberg¹,

Keynan²

1978

J Bacteriol

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The outgrowth of spores of Bacillus subtilis 168 proceeded normally in temperature-sensitive DNA mutants under restrictive conditions and in the absence of DNA synthesis. Two inhibitors of DNA synthesis, nalidixic acid and 6-(phydroxyphenylazo)-uracil, inhibited spore outgrowth under some nutritional conditions; this inhibition of outgrowth however, though not that of DNA synthesis, could be reversed by glucose. The sensitivity of the outgrowing spores to nalidixic acid and 6-(p-hydroxyphenylazo)-uracil inhibition decreased as a function of outgrowth time. The cells became completely resistant to the inhibitors after 90 min. The development of this resistance occurred also in the absence of DNA synthesis. It was concluded that DNA synthesis is not needed for spore outgrowth, and that outgrowing cells and vegetative cells differ in their sensitivity to these inhibitors.

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Resporulation of outgrowing Bacillus subtilis spores

Cited by 20 publications

References 22 publications

Bacterial spores and chemical sporicidal agents.

Bacterial spores and chemical sporicidal agents.

Bacillus subtilis 168 genetic transformation mediated by outgrowing spores: necessity for cell contact

Independence of Bacillus subtilis spore outgrowth from DNA synthesis

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