Difference between revisions of "PMID:21441507"

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'''Salguero, I, Guarino, E and Guzmán, EC'''  (2011) RecA-dependent replication in the nrdA101(Ts) mutant of Escherichia coli under restrictive conditions.''J. Bacteriol.'' '''193''':2851-60
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!align=left  |Abstract
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Cells carrying the thermosensitive nrdA101 allele are able to replicate entire chromosomes at 42°C when new DNA initiation events are inhibited. We investigated the role of the recombination enzymes on the progression of the DNA replication forks in the nrdA101 mutant at 42°C in the presence of rifampin. Using pulsed-field gel electrophoresis (PFGE), we demonstrated that the replication forks stalled and reversed during the replication progression under this restrictive condition. DNA labeling and flow cytometry experiments supported this finding as the deleterious effects found in the RecB-deficient background were suppressed specifically by the absence of RuvABC; however, this did not occur in a RecG-deficient background. Furthermore, we show that the RecA protein is absolutely required for DNA replication in the nrdA101 mutant at restrictive temperature when the replication forks are reversed. The detrimental effect of the recA deletion is not related to the chromosomal degradation caused by the absence of RecA. The inhibition of DNA replication observed in the nrdA101 recA mutant at 42°C in the presence of rifampin was reverted by the presence of the wild-type RecA protein expressed ectopically but only partially suppressed by the RecA protein with an S25P mutation [RecA(S25P)], deficient in the rescue of the stalled replication forks. We propose that RecA is required to maintain the integrity of the reversed forks in the nrdA101 mutant under certain restrictive conditions, supporting the relationship between DNA replication and recombination enzymes through the stabilization and repair of the stalled replication forks.
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[http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=21441507 PubMed]
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Online version:[http://dx.doi.org/10.1128/JB.00109-11 10.1128/JB.00109-11]
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!align=left  |Keywords
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Anti-Bacterial Agents; DNA Replication; DNA, Bacterial; Escherichia coli; Escherichia coli Proteins; Gene Deletion; Genetic Complementation Test; Hot Temperature; Mutant Proteins; Rec A Recombinases; Ribonucleoside Diphosphate Reductase; Rifampin
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==Main Points of the Paper ==
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{{LitSignificance}}
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== Materials and Methods Used ==
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{{LitMaterials}}
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==Phenotype Annotations==
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{|  id="V4f1db665c37fd"  class=" tableEdit Phenotype_Table_2" 
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!|Phenotype of!!Taxon Information!!Genotype Information (if known)!!Condition Information!!OMP ID!!OMP Term Name!!ECO ID!!ECO Term Name!!Notes!!Status
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a single strain under different conditions
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*Taxon: Escherichia coli
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*Strain: K-12
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*Substrain: JS1018
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*NCBI Taxon ID: 83333
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*Genotype of Reference Strain: ''nrdA101RecA''
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*Genotype of Experimental Strain : ''nrdA101recA(del)''
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*Reference Condition: under restrictive temperature conditions
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*Experimental Condition: under restrictive temperature conditions
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For the ''nrdA101''  mutant replication is stalled at restrictive temperatures in the absence of ''recA'' when the replication forks are reversed.
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a mutation or genetic difference within a strain
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*Taxon: Escherichia coli
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*Strain: K-12
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*Substrain: JS1018
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*NCBI Taxon ID: 83333
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*Genotype of Reference Strain: ''nrdA101recB''
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*Genotype of Experimental Strain : ''nrdA101recB(del)''
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*Reference Condition: a missense mutation
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*Experimental Condition: under restrictive temperature conditions
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for the ''nrdA101'' mutant, any negative effects caused by the lack of ''recB'' were negated by RuvABC deficiency.
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==Notes==
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==References==
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<references/>
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[[Category:Publication]]

Latest revision as of 15:36, 23 January 2012

Citation

Salguero, I, Guarino, E and Guzmán, EC (2011) RecA-dependent replication in the nrdA101(Ts) mutant of Escherichia coli under restrictive conditions.J. Bacteriol. 193:2851-60

Abstract

Cells carrying the thermosensitive nrdA101 allele are able to replicate entire chromosomes at 42°C when new DNA initiation events are inhibited. We investigated the role of the recombination enzymes on the progression of the DNA replication forks in the nrdA101 mutant at 42°C in the presence of rifampin. Using pulsed-field gel electrophoresis (PFGE), we demonstrated that the replication forks stalled and reversed during the replication progression under this restrictive condition. DNA labeling and flow cytometry experiments supported this finding as the deleterious effects found in the RecB-deficient background were suppressed specifically by the absence of RuvABC; however, this did not occur in a RecG-deficient background. Furthermore, we show that the RecA protein is absolutely required for DNA replication in the nrdA101 mutant at restrictive temperature when the replication forks are reversed. The detrimental effect of the recA deletion is not related to the chromosomal degradation caused by the absence of RecA. The inhibition of DNA replication observed in the nrdA101 recA mutant at 42°C in the presence of rifampin was reverted by the presence of the wild-type RecA protein expressed ectopically but only partially suppressed by the RecA protein with an S25P mutation [RecA(S25P)], deficient in the rescue of the stalled replication forks. We propose that RecA is required to maintain the integrity of the reversed forks in the nrdA101 mutant under certain restrictive conditions, supporting the relationship between DNA replication and recombination enzymes through the stabilization and repair of the stalled replication forks.

Links

PubMed Online version:10.1128/JB.00109-11

Keywords

Anti-Bacterial Agents; DNA Replication; DNA, Bacterial; Escherichia coli; Escherichia coli Proteins; Gene Deletion; Genetic Complementation Test; Hot Temperature; Mutant Proteins; Rec A Recombinases; Ribonucleoside Diphosphate Reductase; Rifampin

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Main Points of the Paper

Please summarize the main points of the paper.

Materials and Methods Used

Please list the materials and methods used in this paper (strains, plasmids, antibodies, etc).

Phenotype Annotations

See Help:AnnotationTable for details on how to edit this table.
<protect>

Phenotype of Taxon Information Genotype Information (if known) Condition Information OMP ID OMP Term Name ECO ID ECO Term Name Notes Status

a single strain under different conditions

  • Taxon: Escherichia coli
  • Strain: K-12
  • Substrain: JS1018
  • NCBI Taxon ID: 83333
  • Genotype of Reference Strain: nrdA101RecA
  • Genotype of Experimental Strain : nrdA101recA(del)
  • Reference Condition: under restrictive temperature conditions
  • Experimental Condition: under restrictive temperature conditions

For the nrdA101 mutant replication is stalled at restrictive temperatures in the absence of recA when the replication forks are reversed.

a mutation or genetic difference within a strain

  • Taxon: Escherichia coli
  • Strain: K-12
  • Substrain: JS1018
  • NCBI Taxon ID: 83333
  • Genotype of Reference Strain: nrdA101recB
  • Genotype of Experimental Strain : nrdA101recB(del)
  • Reference Condition: a missense mutation
  • Experimental Condition: under restrictive temperature conditions

for the nrdA101 mutant, any negative effects caused by the lack of recB were negated by RuvABC deficiency.


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</protect>

Notes

References

See Help:References for how to manage references in omp dev.

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