ArticleDNA repair2012
Simple and efficient purification of Escherichia coli DNA polymerase V: cofactor requirements for optimal activity and processivity in vitro.
Article in DNA repair, 2012. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 30 papers.
What it found
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The abstract states no effect estimate the extractor could read, or names no intervention and outcome on the map, so this paper lights no cell and moves no belief. It is still indexed, cited and linked below.
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Who cites it
30 citing papers in PubMed, 39 citations in OpenAlex.
- Acinetobacter baumannii encodes multiple mutagenically and biochemically active DNA polymerase V variants.Nucleic acids research · 2026Article
- Pathogen-encoded Rum DNA polymerase drives rapid bacterial drug resistance.Nucleic acids research · 2024Article
- Escherichia coli DNA replication: the old model organism still holds many surprises.FEMS microbiology reviews · 2024Review
- E3 ubiquitin ligase RNF2 protects polymerase ι from destabilization.Biochimica et biophysica acta. Molecular cell research · 2024Article
- A Novel Interaction Between RAD23A/B and Y-family DNA Polymerases.Journal of molecular biology · 2023Article
- Integration of molecular modelling andFrontiers in cellular and infection microbiology · 2023Article
- During Translesion Synthesis, Escherichia coli DinB89 (T120P) Alters Interactions of DinB (Pol IV) with Pol III Subunit Assemblies and SSB, but Not with the β Clamp.Journal of bacteriology · 2022Article
- Article
- Role of RNase H enzymes in maintaining genome stability in Escherichia coli expressing a steric-gate mutant of pol VDNA repair · 2019Article
- The SOS system: A complex and tightly regulated response to DNA damage.Environmental and molecular mutagenesis · 2019Review
- DNA polymerase ι is acetylated in response to SScientific reports · 2019Article
- Conformational regulation of Escherichia coli DNA polymerase V by RecA and ATP.PLoS genetics · 2019Article
- The recombination mediator proteins RecFOR maintain RecA* levels for maximal DNA polymerase V Mut activity.The Journal of biological chemistry · 2019Article
- SetRDNA repair · 2019Article
- Purification and interactions of the MucA' and MucB proteins constituting the DNA polymerase RI.Genes and environment : the official journal of the Japanese Environmental Mutagen Society · 2019Article
- Ribonucleotide discrimination by translesion synthesis DNA polymerases.Critical reviews in biochemistry and molecular biology · 2018Review
- Insights into the complex levels of regulation imposed on Escherichia coli DNA polymerase V.DNA repair · 2016Review
- Exchange between Escherichia coli polymerases II and III on a processivity clamp.Nucleic acids research · 2016Article
- Molecular architecture of the Ub-PCNA/Pol η complex bound to DNA.Scientific reports · 2015Article
- Regulation of Mutagenic DNA Polymerase V Activation in Space and Time.PLoS genetics · 2015Article
Corrections and comments
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Authors and funding
4 authors at 2 institutions in 1 country.
Funding
Abstract
Most damage induced mutagenesis in Escherichia coli is dependent upon the UmuD'(2)C protein complex, which comprises DNA polymerase V (pol V). Biochemical characterization of pol V has been hindered by the fact that the enzyme is notoriously difficult to purify, largely because overproduced UmuC is insoluble. Here, we report a simple and efficient protocol for the rapid purification of milligram quantities of pol V from just 4 L of bacterial culture. Rather than over producing the UmuC protein, it was expressed at low basal levels, while UmuD'(2)C was expressed in trans from a high copy-number plasmid with an inducible promoter. We have also developed strategies to purify the β-clamp and γ-clamp loader free from contaminating polymerases. Using these highly purified proteins, we determined the cofactor requirements for optimal activity of pol V in vitro and found that pol V shows robust activity on an SSB-coated circular DNA template in the presence of the β/γ-complex and a RecA nucleoprotein filament (RecA*) formed in trans. This strong activity was attributed to the unexpectedly high processivity of pol V Mut (UmuD'(2)C · RecA · ATP), which was efficiently recruited to a primer terminus by SSB.
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Registered trials
Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.