The role of DNA polymerase I, II and III in the replication of the bacteriocinogenic factor Clo DF13

Veltkamp, E.; Nijkamp, H. J. J.

doi:10.1007/bf00276588

Cited by 25 publications

(20 citation statements)

References 25 publications

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“…We therefore performed experiments to estimate the number of Clo DF13 copies in wild-type and mutant cloacinogenic cells by the method of Durkacz and Sherratt (4). The Clo DF13 plasmid requires DNA polymerase I for its replication (23). Strain TS214(Clo DF13) [=N3165] is temperature sensitive for DNA polymerase I; the Clo DF13 plasmid is unable to replicate at the nonpermissive temperature of 43 C. Cell divisions of strain TS214(Clo DF13) occurs at 43 C and results in the generation of plasmidfree cells.…”

Section: Resultsmentioning

confidence: 99%

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Isolation and Characterization of a Copy Mutant of the Bacteriocinogenic Plasmid Clo DF13

Kool

Nijkamp

1974

J Bacteriol

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After nitrosoguanidine mutagenesis, strain Escherichia coli P678-54, bacteriocinogenic for Clo DF13, yielded a mutant strain that showed an enhanced bacteriocin production. The results from conjugation experiments indicated that the mutation, responsible for the enhanced bacteriocin production, is located on the Clo DF13 plasmid. The following properties of strains harboring the mutant Clo DF13 plasmid could be observed. (i) The bacteriocin production in these strains can be further enhanced at least fourfold by mitomycin C. (ii) The fraction of spontaneously induced cells, as revealed by lacunae experiments, in cultures of these strains is about nine times higher than in cultures of wild-type Clo DF13-harboring strains. (iii) Chromosomeless minicells from strain P678-54 harboring the mutant Clo DF13 plasmid synthesize about six times more deoxyribonucleic acid, ribonucleic acid, and protein as compared to wild-type Clo DF13-harboring minicells. (iv) Analysis of this mutant Clo DF13-specific ribonucleic acid and protein on polyacrylamide gels revealed mainly the same ribonucleic acid and polypeptide species as synthesized by the wild-type Clo DF13 minicells, but in larger amounts (Kool et al., 1974). (v) Segregation experiments, using a strain with temperature-sensitive polymerase I, show that mutant Clo DF13-harboring cells contain an average of 70 Clo DF13 copies per cell, whereas wild-type Clo DF13-harboring cells contain only about 10 Clo DF13 copies per cell. The data presented in this paper indicate that the mutation on the Clo DF13 plasmid leads to an altered control of Clo DF13 replication and results in an enhanced number of Clo DF13 copies per cell. As a secondary effect, this enhanced number of Clo DF13 copies enhances the probability of "spontaneous" induction per cell. Since the mutation is plasmid specific and affects the number of plasmid copies produced, one can conclude that the Clo DF13 plasmid is not dependent solely on chromosomal information, but that at least plasmid base sequences are involved in Clo DF13 plasmid replication. 569 on August 5, 2020 by guest

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Section: Resultsmentioning

confidence: 99%

“…Therefore, the observed sixfold-increased RNA and protein synthesis in mutant cloacinogenic minicells may be due to increased gene dosage. The is known to inhibit Clo DF13 replication in minicells (23). Symbols: 0, incorporation by minicells isolated from P678-54(Clo DF13) A, incorporation by those isolated from P678-54(Clo DF13-M3).…”

Section: Discussionmentioning

confidence: 99%

Isolation and Characterization of a Copy Mutant of the Bacteriocinogenic Plasmid Clo DF13

Kool

Nijkamp

1974

J Bacteriol

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“…For the construction of deletion mutants, pJN67 DNA was digested with Hpa I in the Hpa I incubation mixture. Sub-HPalal r 4 1 pJN 67…”

Section: Mapping Of Tn901 Insertions On Clo Df13mentioning

confidence: 99%

“…Plasmid Clo DF13, with a molecular weight of 5.75 MegaDaltons (MD), has been extensively used in our laboratory to study the replication, gene function and genetic organisation of bacterial plasmids (1)(2)(3)(4)(5)(6)(7). It was observed that this plasmid, which is present in Escherichia coli to the extend of about 10 copies per cell, directs the synthesis of at least four m-RNA's and eight proteins (2,6).…”

Section: Introductionmentioning

confidence: 99%

In vitro construction of deletion mutants of the bacteriocinogenic plasmid Clo DF13

et al. 1978

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The isolation and characterization of deletion mutants of the bacteriocinogenic plasmid Clo DF13 is described. To construct these deletion mutants, DNA of Clo DF13::Tn901 and Clo DF13-rep3::Tn901 plasmids was digested with restriction endonucleases, ligated with T4 ligase and introduced by transformation into Escherichia coli. The presence of the ampicilline transposon Tn901 facilitated the selection of plasmids. The resulting Clo DF13::Tn901 deletion mutants were analyzed by digestion with restriction endonucleases and electron microscopy. From the properties of the various deletion mutants it was concluded that a Clo DF13 DNA region, extending from 5 to 11.5% on the physical map, is essential for the replication of Clo DF13. This region, comprising about 600 base pairs, contains in addition to an origin of replication, DNA sequences which are involved in the regulation of Clo DF13 DNA replication. Furthermore it was observed that in case of the Clo DF13 copy mutant, Clo DF13-rep3, deletion of the 43% to 63% part of the plasmid genome, resulted in the generation of multimeric plasmid structures, accompanied with an impaired segregation of the plasmids to daughter cells.

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“…The small nonconjugative plasmid CloDF13, is present in Escherichia coli to an extent of about 10 copies per cell (36). Copy control mutants of CloDF13 have been isolated (3, 13), such as CloDF13 cop-3 with a plasmid copy number of about 70 (37).…”

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Transcription of bacteriocinogenic plasmid CloDF13 in vivo and in vitro: structure of the cloacin immunity operon

Elzen¹,

Konings²,

Veltkamp³

et al. 1980

J Bacteriol

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Escherichia coli minicells harboring plasmid CloDF13 synthesized at least 25 messenger ribonucleic acid (RNA) species; three of these RNAs, a 2,400-, a 2,200-, and a 100-nucleotide RNA, were synthesized in relatively large amounts. Using insertion and deletion mutants of CloDF13 as well as an RNA blotting technique, we could demonstrate that these three RNAs are transcripts from the CloDF13 DNA region from 0 to 40%. This region contains the cloacin and immunity genes and the genetic information involved in plasmid DNA replication. A transcription map of this region is presented and discussed. The data indicate that the cloacin and immunity genes were coordinately transcribed into messenger RNAs of about 2,400 and 2,200 nucleotides, which differ in length at their 3' terminus. RNA polymerase binding studies and in vitro transcription assays indicated that transcription of these genes initiates at a promoter located around 32% on the CloDF13 map. Furthermore, it is shown that a 100-nucleotide RNA is encoded by the CloDF13 DNA region between 7.7 and 8.8% on the plasmid genome; the synthesis of this RNA proceeds in a direction opposite to the transcription of the cloacin and immunity genes.tively labeled DNA. Our results indicate that 579 on August 5, 2020 by guest

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The role of DNA polymerase I, II and III in the replication of the bacteriocinogenic factor Clo DF13

Cited by 25 publications

References 25 publications

Isolation and Characterization of a Copy Mutant of the Bacteriocinogenic Plasmid Clo DF13

Isolation and Characterization of a Copy Mutant of the Bacteriocinogenic Plasmid Clo DF13

In vitro construction of deletion mutants of the bacteriocinogenic plasmid Clo DF13

Transcription of bacteriocinogenic plasmid CloDF13 in vivo and in vitro: structure of the cloacin immunity operon

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