Properties of four mutants of Escherichia coli defective in genetic recombination

Putte, Pieter van de; Zwenk, H.; Rörsch, A.

doi:10.1016/0027-5107(66)90048-0

Cited by 86 publications

(32 citation statements)

References 8 publications

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“…3B, ⌬recA, lanes a-c). This was expected, because neither phage replication nor its recombination in freely replicating crosses are dependent on the general recombination functions of the host (Takano 1966;van de Putte et al 1966;Brooks and Clark 1967). When phages were cut, but not repressed, some DNA replication still occurred, but the amount of the double-deletion recombinant was lower than it was in recA + cells (Fig.…”

Section: Double-strand Ends Stimulate Recombinationmentioning

confidence: 99%

Double-strand end repair via the RecBC pathway in Escherichia coli primes DNA replication

Kuzminov¹,

Stahl²

1999

Genes & Development

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To study the relationship between homologous recombination and DNA replication in Escherichia coli, we monitored the behavior of phage chromosomes, repressed or not for gene activities. Recombination in our system is stimulated both by DNA replication and by experimentally introduced double-strand ends, supporting the idea that DNA replication generates occasional double-strand ends. We report that the RecBC recombinational pathway of E. coli uses double-strand ends to prime DNA synthesis, implying a circular relationship between DNA replication and recombination and suggesting that the primary role of recombination is in the repair of disintegrated replication forks arising during vegetative reproduction.

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Section: Double-strand Ends Stimulate Recombinationmentioning

confidence: 99%

Double-strand end repair via the RecBC pathway in Escherichia coli primes DNA replication

Kuzminov¹,

Stahl²

1999

Genes & Development

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“…Recombinationdeficient mutants of bacteriophage and bacteria have been useful in the study of recombination processes (1)(2)(3)(4)(5)(6)(7)(8)(9)(10). Recombination deficient, radiation-sensitive mutants of Bacillus subtilis have been isolated and described by several investigators (11)(12)(13)(14).…”

mentioning

confidence: 99%

Fate of Transforming DNA After Uptake by Competent Bacillus subtilis : Failure of Donor DNA to Replicate in a Recombination-Deficient Recipient

Davidoff-Abelson¹,

Dubnau²

1971

Proc. Natl. Acad. Sci. U.S.A.

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The fate of radioactively-labeled transforming DNA was studied in a recombination-deficient strain of Bacillus subtilis that carried the recB2 mutation (Rec-) Bacterial transformation is an advantageous system for the study of recombination, since the fate of purified donor DNA can be studied in the course of the different steps before, during, and after integration. Recombinationdeficient mutants of bacteriophage and bacteria have been useful in the study of recombination processes (1-10). Recombination deficient, radiation-sensitive mutants of Bacillus subtilis have been isolated and described by several investigators (11)(12)(13)(14). This paper reports the results of experiments in which the fate of transforming DNA was examined in one of these mutants (recB2). Our results indicate that in recB2, the donor-recipient complex is formed to the same extent as in an isogenic Rec+ strain, but that donor DNA fails to replicate in the recipient after formation of the complex. MATERIALS AND METHODS Bacterial strainsThe bacterial strains used were derivatives of Bacillus subtilis 168. Strains BD170 (thr-5 trp-2) and BD191 (thr-5 trp-2 recB2) were derived from the same parent and were isogenic for all markers except the recB2 marker, which was introduced by transformation as was described (15). The recB2 marker is derived from the mitomycin-sensitive strain mc-1 (12). RecB2 has been shown to be radiation sensitive and deficient in both transformability and transducibility (12)(13)(14). Strain BD204 (hisB2 thy) was used to prepare radioactively-labeled transforming DNA.BD55 (trp-2 hisBW), known also as SB25, was used to assay the biological activity of DNA fractions isolated from the transformed cells. The trp-2 and hisB2 markers are cotransformed at a frequency of about 60%. Media and solutionsThe media for plating and for the development of competence were prepared as described (16). Transformed cells were washed in SP buffer (0.15 M NaCl-0.02 M K2HPO4, pH 7.5). The washed samples were resuspended in lysis medium (0.05 M NaCl-0.1 M EDTA, pH 6.9).SPII medium contained Spizizen salts (17), 0.02% casamino acids, 0.1% Difco yeast extract, 0.5% glucose, 0.5 mM CaC12, 2.5 mM MgCl2, and 50,gg/ml of each required amino acid. Re-extraction experimentsCompetent cells were prepared and frozen as was described (16). Frozen competent Rec+ and Rec-cells were quickly thawed and incubated for 5 min at 370C. To begin the reaction, [3H]DNA, isolated from BD204 as described (16), was then added to a final concentration of 1-2 gg/ml. The specific activity of the DNA preparations ranged from 4 to 6 X 105 cpm/htg. The mixture was incubated at 370C for 7 min or 9 min, at which time an excess (100 ,Ag/ml) of salmonsperm DNA (Calbiochem) was added to stop further DNA uptake. In some experiments the reaction mixture was then transferred to 300C. At different times after addition of the salmon-sperm DNA, samples of the transformed cells were chilled rapidly, washed three times by centrifugation in icecold SP, and resuspended in lysis ...

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“…Recombinational repair is another form of repair which deals with ultraviolet-induced lesions in E. coli, though much less is known about its mechanism of action. Recombinational repair (9,(16)(17)(18)(19) has been proposed to act by removing lesions which are too close to the replication fork to be dealt with by repair synthesis (excision repair) (20). Strand excision too near the replication fork might very probably lead to lethal damage equivalent to a double-stranded break in DNA.…”

Section: Other Types Of Repairmentioning

confidence: 99%

“…It has not been definitively demonstrated whether mammalian cells engage in recombinational repair, and in fact, the main evidence in E. coli stems from the observation that this form of repair is governed by the locus controlling recombination (16,17). In addition, there is biochemical evidence supporting (18) the view that recombinational events provide a means of dealing with genetic lesions but this evidence is less convincing than that available on repair-synthesis.…”

Section: Other Types Of Repairmentioning

confidence: 99%

The role of repair in environmental mutagenesis.

Flamm¹

1973

Environ Health Perspect

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Properties of four mutants of Escherichia coli defective in genetic recombination

Cited by 86 publications

References 8 publications

Double-strand end repair via the RecBC pathway in Escherichia coli primes DNA replication

Double-strand end repair via the RecBC pathway in Escherichia coli primes DNA replication

Fate of Transforming DNA After Uptake by Competent Bacillus subtilis : Failure of Donor DNA to Replicate in a Recombination-Deficient Recipient

The role of repair in environmental mutagenesis.

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