On the mechanism of conjugation in Escherichia coli K12

Бреслер, С. Е.; Lanzov, Vladislav A.; Likhachev, V T

doi:10.1007/bf00267240

Cited by 13 publications

(14 citation statements)

References 11 publications

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“…On the other hand, previous reports addressing the possible role of extensive DNA replication in homologous recombination in E. coli are conflicting. The yield of recombinants after conjugation in a dnaB(Ts) recipient at the restrictive temperature was reported to be either greatly decreased (9) or stimulated (15). The recombination-by-replication model (Fig.…”

Section: Discussionmentioning

confidence: 99%

The DNA replication priming protein, PriA, is required for homologous recombination and double-strand break repair

et al. 1996

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The PriA protein, a component of the X174-type primosome, was previously shown to be essential for damage-inducible DNA replication in Escherichia coli, termed inducible stable DNA replication. Here, we show that priA::kan null mutants are defective in transductional and conjugational homologous recombination and are hypersensitive to mitomycin C and gamma rays, which cause double-strand breaks. The introduction of a plasmid carrying the priA300 allele, which encodes a mutant PriA protein capable of catalyzing the assembly of an active primosome but which is missing the n-pas-dependent ATPase, helicase, and translocase activities associated with PriA, alleviates the defects of priA::kan mutants in homologous recombination, double-strand break repair, and inducible stable DNA replication. Furthermore, spa-47, which was isolated as a suppressor of the broth sensitivity of priA::kan mutants, suppresses the Rec ؊ and mitomycin C sensitivity phenotypes of priA::kan mutants. The spa-47 suppressor mutation maps within or very near dnaC. These results suggest that PriA-dependent primosome assembly is crucial for both homologous recombination and double-strand break repair and support the proposal that these processes in E. coli involve extensive DNA replication.Homologous recombination, a ubiquitous activity in both prokaryotes and eukaryotes, not only is a means by which to generate genetic diversity but also plays a crucial role in the repair of DNA damage, including double-strand breaks (DSBs) (8,12). Despite recent advances in our understanding of the mechanism of homologous recombination, the extent to which DNA synthesis might be involved in the process is largely unknown. PriA, a component of the priming system which primes DNA synthesis in the initiation of X174 phage and ColE1-type plasmid DNA replication, has recently been shown to play an essential role in the initiation of DNA damage-inducible chromosome replication in Escherichia coli, termed inducible stable DNA replication (iSDR) (25). Here, we show that priA null mutants are defective in homologous recombination and are hypersensitive to chemical and physical agents that cause DSBs. These results strongly support the notion that extensive DNA replication is involved in homologous recombination and DSB repair.The X174-type primosome, originally discovered in the study of the initiation of phage X174 DNA replication, consists of several E. coli proteins (see reference 23 for a review

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Section: Discussionmentioning

confidence: 99%

The DNA replication priming protein, PriA, is required for homologous recombination and double-strand break repair

et al. 1996

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“…This observation had permitted us to measure conjugal replication during such a mating and to correlate this synthesis with transfer of DNA to the recipient minicells. Bresler et al (5) first noted conjugal replication when vegetative DNA synthesis was inhibited by temperature in a study using Hfr donor cells. Marinus and Adelberg (27) and Vapnek and Rupp (44) found this type of replication during the transfer of F'lac and F, respectively.…”

Section: Discussionmentioning

confidence: 99%

“…We first determined whether recipient minicells, like recipient cells (5,27,44), could stimulate incorporation of [3H]thymidine by dnaB(ts) donor cells at temperatures restrictive for vegetative DNA replication. At least 12 times more ['Hithymidine was incorporated into acid-insoluble material when x1284 [R64-11+ dnaB(ts) ] donor cells were incubated for 30 min at 42 C in the presence of minicells derived from a Fparent strain (X925) than when they were incubated-alone ( Fig.…”

Section: Stimulation Of Conjugal Dna Replicationmentioning

confidence: 99%

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Conjugal Deoxyribonucleic Acid Replication by Escherichia coli K-12: Stimulation in dnaB (ts) Donors by Minicells

Fenwick

Curtiss

1973

J Bacteriol

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R64-11+ donor cells that are thermosensitive for vegetative DNA replication will synthesize DNA at the restrictive temperature when recipient minicells are present. This is conjugal DNA replication because it is R64-11 DNA that is being synthesized. and there is no DNA synthesis if minicells that cannot be recipients of R64-11 DNA are used. The plasmid DNA present in the donor cells before mating is transferred to recipient minicells within the first 20 min of mating, but additional copies of plasmid DNA synthesized during the mating continue to be transferred for at least 90 min. However, the transfer of R64-11 DNA to minicells is not continuous because the plasmid DNA in minicells is the size of one R64-11 molecule or smaller, and there are delays between the rounds of plasmid transfer. DNA is synthesized in minicells during conjugation, but this DNA has a molecular weight much smaller than that of R64-11. Thus, recipient minicells are defective and are not able to complete the synthesis of a DNA strand complementary to the single-stranded R64-11 DNA received from the donor cell. 1212 on July 31, 2020 by guest

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“…He isolated donor and recipient strains that were resistant to the drug and found that transfer of F'lac was only inhibited when the donor cells were sensitive. These findings were extended to conjugal transfer of DNA from Hfr (7,12,16) and F+ (9) donor cells. Hollom and Pritchard (21) originally described the effect of nalidixic acid on conjugation as reversible.…”

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confidence: 87%

Conjugal Deoxyribonucleic Acid Replication by Escherichia coli K-12: Effect of Nalidixic Acid

Fenwick

Curtiss

1973

J Bacteriol

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During the conjugal transfer of the R64-11 plasmid at 42 C from donor cells thermosensitive for vegetative deoxyribonucleic acid (DNA) synthesis to recipient minicells, the plasmids are conjugally replicated in the donor cells. This conjugal replication is inhibited by nalidixic acid, and the degree of inhibition is comparable to the reduction in the amount of plasmid DNA transferred to the recipient minicells in the presence of the drug. In addition, the size of DNA transferred to the minicells and the fraction of conjugally replicated DNA in the donor cells that can be isolated as closed-circular plasmid DNA under alkaline conditions are both reduced by nalidixic acid. When the drug is added to a mating that is underway, the rate of conjugal replication is immediately reduced. This change is accompanied by a reduction in the amount of conjugally replicated DNA in the donor cells that can be isolated as closed-circular plasmid DNA. Furthermore, conjugally replicated plasmid DNA that is not associated with the donor cell membrane becomes membrane bound after the addition of nalidixic acid.

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On the mechanism of conjugation in Escherichia coli K12

Cited by 13 publications

References 11 publications

The DNA replication priming protein, PriA, is required for homologous recombination and double-strand break repair

The DNA replication priming protein, PriA, is required for homologous recombination and double-strand break repair

Conjugal Deoxyribonucleic Acid Replication by Escherichia coli K-12: Stimulation in dnaB (ts) Donors by Minicells

Conjugal Deoxyribonucleic Acid Replication by Escherichia coli K-12: Effect of Nalidixic Acid

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