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PMID: 10430924 Published · ppublish English Comparative Study Journal Article Research Support, U.S. Gov't, P.H.S.

A phenotype for enigmatic DNA polymerase II: a pivotal role for pol II in replication restart in UV-irradiated Escherichia coli.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 96 ·No. 16 ·1999-08-03 ·Pages 9224-9

Rangarajan S, Woodgate R, Goodman MF

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.

MeSH Terms
Bacterial Proteins/genetics DNA Damage DNA Polymerase II/genetics,metabolism DNA Polymerase III/metabolism DNA Replication/genetics,radiation effects DNA, Bacterial/biosynthesis,radiation effects DNA-Directed DNA Polymerase Dose-Response Relationship, Radiation Escherichia coli/enzymology,genetics,radiation effects Escherichia coli Proteins Genotype Phenotype Polymerase Chain Reaction SOS Response, Genetics/genetics,radiation effects Time Factors Ultraviolet Rays
Chemicals
Bacterial Proteins DNA, Bacterial Escherichia coli Proteins UmuC protein, E coli DNA Polymerase II DNA Polymerase III DNA-Directed DNA Polymerase UmuD protein, E coli
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Rangarajan S
Department of Biological Sciences, Hedco Molecular Biology Laboratories, University of Southern California, Los Angeles, CA 90089-1340, USA.
Woodgate R
Goodman M F
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Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
DOI
10.1073/pnas.96.16.9224
Published
1999-08-03
Pages
9224-9
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC17761
Subset
IM
Grants
NIGMS NIH HHS · GM42554 · United States
External Links
PubMed source DOI: 10.1073/pnas.96.16.9224 PMC Full Text View journal details
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