Difference between revisions of "PMID:10430924"

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Latest revision as of 16:04, 21 March 2012

Citation

Rangarajan, S, Woodgate, R and Goodman, MF (1999) A phenotype for enigmatic DNA polymerase II: a pivotal role for pol II in replication restart in UV-irradiated Escherichia coli.Proc. Natl. Acad. Sci. U.S.A. 96:9224-9

Abstract

DNA synthesis in Escherichia coli is inhibited transiently after UV irradiation. Induced replisome reactivation or "replication restart" occurs shortly thereafter, allowing cells to complete replication of damaged genomes. At the present time, the molecular mechanism underlying replication restart is not understood. DNA polymerase II (pol II), encoded by the dinA (polB) gene, is induced as part of the global SOS response to DNA damage. Here we show that pol II plays a pivotal role in resuming DNA replication in cells exposed to UV irradiation. There is a 50-min delay in replication restart in mutant cells lacking pol II. Although replication restart appears normal in DeltaumuDC strains containing pol II, the restart process is delayed for >90 min in cells lacking both pol II and UmuD'(2)C. Because of the presence of pol II, a transient replication-restart burst is observed in a "quick-stop" temperature-sensitive pol III mutant (dnaE486) at nonpermissive temperature. However, complete recovery of DNA synthesis requires the concerted action of both pol II and pol III. Our data demonstrate that pol II and UmuD'(2)C act in independent pathways of replication restart, thereby providing a phenotype for pol II in the repair of UV-damaged DNA.

Links

PubMed

Keywords

Bacterial Proteins; DNA Damage; DNA Polymerase II; DNA Polymerase III; DNA Replication; DNA, Bacterial; DNA-Directed DNA Polymerase; Dose-Response Relationship, Radiation; Escherichia coli; Escherichia coli Proteins; Genotype; Phenotype; Polymerase Chain Reaction; SOS Response (Genetics); Time Factors; Ultraviolet Rays

Main Points of the Paper

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Materials and Methods Used

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Phenotype Annotations

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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

a mutation or genetic difference within a strain

  • Taxon: Escherichia coli
  • Strain: K-12
  • Substrain: SH2101
  • NCBI Taxon ID: 83333
  • Genotype of Reference Strain: 'araD-polB+
  • Genotype of Experimental Strain : (araD-polB)(del)::omega
  • Reference Condition:
ECO:0000004

cell fractionation data

After the insertion of cat there was a significant delay in the resumption of DNA replication after exposure to UV radiation. This is in comparison to the wild-type strain. See figure 2.

a mutation or genetic difference within a strain

  • Taxon: Escherichia coli
  • Strain: K-12
  • Substrain: RW622
  • NCBI Taxon ID: 83333
  • Genotype of Reference Strain: dnaE486 araD-polB +
  • Genotype of Experimental Strain : dnaE786 (araD-polB)(del)::omega
  • Reference Condition:
ECO:0000004

cell fractionation data

At a permissive temperature, 30oC, there was a significant delay in the resumption of DNA replication. See Figure 4B.

a mutation or genetic difference within a strain

  • Taxon: Escherichia coli
  • Strain: K-12
  • Substrain: SR1157U
  • NCBI Taxon ID: 83333
  • Genotype of Reference Strain: polB+ umuDC+
  • Genotype of Experimental Strain : (araD-polB)(del)::omega "umuDC(del)::cat
  • Reference Condition:
ECO:0000004

cell fractionation data

Sensitivity toward UV radiation was increased 2 to 3 fold in the double mutant. See figure 1.

a mutation or genetic difference within a strain

  • Taxon: Escherichia coli
  • Strain: K-12
  • Substrain: SR1157U
  • NCBI Taxon ID: 83333
  • Genotype of Reference Strain: polB+ umuDC+
  • Genotype of Experimental Strain : (araD-polB)(del)::omega "umuDC(del)::cat
  • Reference Condition:
ECO:0000004

cell fractionation data

There was a significant delay in the resumption of DNA replication after exposure to UV radiation. See figure 1.

a mutation or genetic difference within a strain

  • Taxon: Escherichia coli
  • Strain: K-12
  • Substrain: RW82
  • NCBI Taxon ID: 83333
  • Genotype of Reference Strain: umuDC+
  • Genotype of Experimental Strain : umuDC595(del)::cat
  • Reference Condition:
ECO:0000004

cell fractionation data

After the insertion of the chloramephenicol acetylransferase gene (cat) there was a moderate increase in sensitivity toward UV radiation. See figure 1.

a mutation or genetic difference within a strain

  • Taxon: Escherichia coli
  • Strain: K-12
  • Substrain: RW82
  • NCBI Taxon ID: 83333
  • Genotype of Reference Strain: umuDC+
  • Genotype of Experimental Strain : umuDC595(del)::cat
  • Reference Condition:
ECO:0000004

cell fractionation data

After the insertion of the chloramephenicol acetylransferase gene (cat) there was a slight delay in the resumption of DNA replication after exposure to UV radiation.


Notes

References

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