Single-molecule live-cell imaging reveals RecB-dependent function of DNA polymerase IV in double strand break repair.

Nucleic Acids Research
S. S. HenrikusAndrew Robinson

Abstract

Several functions have been proposed for the Escherichia coli DNA polymerase IV (pol IV). Although much research has focused on a potential role for pol IV in assisting pol III replisomes in the bypass of lesions, pol IV is rarely found at the replication fork in vivo. Pol IV is expressed at increased levels in E. coli cells exposed to exogenous DNA damaging agents, including many commonly used antibiotics. Here we present live-cell single-molecule microscopy measurements indicating that double-strand breaks induced by antibiotics strongly stimulate pol IV activity. Exposure to the antibiotics ciprofloxacin and trimethoprim leads to the formation of double strand breaks in E. coli cells. RecA and pol IV foci increase after treatment and exhibit strong colocalization. The induction of the SOS response, the appearance of RecA foci, the appearance of pol IV foci and RecA-pol IV colocalization are all dependent on RecB function. The positioning of pol IV foci likely reflects a physical interaction with the RecA* nucleoprotein filaments that has been detected previously in vitro. Our observations provide an in vivo substantiation of a direct role for pol IV in double strand break repair in cells treated with double strand break-indu...Continue Reading

References

Oct 1, 1990·Antonie van Leeuwenhoek·G StorzB N Ames
May 1, 1985·Proceedings of the National Academy of Sciences of the United States of America·D G EnnisD W Mount
Mar 1, 1988·Proceedings of the National Academy of Sciences of the United States of America·S E BurckhardtH Echols
Jan 1, 1983·Methods in Enzymology·J Messing
Sep 1, 1993·Proceedings of the National Academy of Sciences of the United States of America·E G FrankR Woodgate
Sep 17, 1996·Proceedings of the National Academy of Sciences of the United States of America·E G FrankR Woodgate
Sep 5, 1997·Science·F R BlattnerY Shao
Dec 10, 1999·Microbiology and Molecular Biology Reviews : MMBR·A Kuzminov
Mar 20, 2001·Nature Reviews. Molecular Cell Biology·M F Goodman, B Tippin
Oct 9, 2002·Journal of Molecular Biology·Jennifer L LavrrarMark A McIntosh
Feb 25, 2003·The Journal of Biological Chemistry·Shelley L LusettiMichael M Cox
Oct 8, 2003·The Journal of Biological Chemistry·Nami HarutaMichael M Cox
May 5, 2005·PLoS Biology·Ryan T CirzFloyd E Romesberg
Mar 23, 2007·Cell Cycle·Daniel F JaroszGraham C Walker
Oct 6, 2007·Critical Reviews in Biochemistry and Molecular Biology·Patricia L Foster
Dec 27, 2007·Molecular Cell·Veronica G GodoyGraham C Walker
Jun 20, 2008·Proceedings of the National Academy of Sciences of the United States of America·Bifeng YuanYinsheng Wang
Aug 14, 2008·The Journal of Biological Chemistry·Anuradha KumariR Stephen Lloyd
Nov 11, 2008·Journal of Bacteriology·Susan E CohenGraham C Walker
Dec 5, 2008·Microbiology and Molecular Biology Reviews : MMBR·Mark S Dillingham, Stephen C Kowalczykowski
Jun 9, 2009·Journal of Bacteriology·Dennis R HarrisJohn R Battista
Sep 15, 2009·Current Topics in Medicinal Chemistry·Karl DrlicaXilin Zhao
Aug 11, 2010·Proceedings of the National Academy of Sciences of the United States of America·Susan E CohenGraham C Walker
Feb 18, 2011·Transcription·Susan E Cohen, Graham C Walker
Mar 19, 2011·Molecular Microbiology·Oleksii SliusarenkoChristine Jacobs-Wagner
May 31, 2011·Frontiers in Bioscience (Landmark Edition)·Jason M WalshPenny J Beuning
Jul 22, 2011·Nature·Gene-Wei Li, X Sunney Xie

❮ Previous
Next ❯

Citations

Nov 17, 2020·Frontiers in Microbiology·Sarah A Revitt-Mills, Andrew Robinson
May 1, 2021·Biology·Camille Henry, Sarah S Henrikus
Jul 10, 2021·Journal of Molecular Biology·Claudia DanilowiczMara Prentiss

❮ Previous
Next ❯

Methods Mentioned

BETA
fluorescence microscopy
PCR
gel-filtration
chips
chip
Fluorescence

Software Mentioned

MATLAB
Peak Fitter
MicrobeTracker

Related Concepts

Trending Feeds

COVID-19

Coronaviruses encompass a large family of viruses that cause the common cold as well as more serious diseases, such as the ongoing outbreak of coronavirus disease 2019 (COVID-19; formally known as 2019-nCoV). Coronaviruses can spread from animals to humans; symptoms include fever, cough, shortness of breath, and breathing difficulties; in more severe cases, infection can lead to death. This feed covers recent research on COVID-19.

Blastomycosis

Blastomycosis fungal infections spread through inhaling Blastomyces dermatitidis spores. Discover the latest research on blastomycosis fungal infections here.

Nuclear Pore Complex in ALS/FTD

Alterations in nucleocytoplasmic transport, controlled by the nuclear pore complex, may be involved in the pathomechanism underlying multiple neurodegenerative diseases including Amyotrophic Lateral Sclerosis and Frontotemporal Dementia. Here is the latest research on the nuclear pore complex in ALS and FTD.

Applications of Molecular Barcoding

The concept of molecular barcoding is that each original DNA or RNA molecule is attached to a unique sequence barcode. Sequence reads having different barcodes represent different original molecules, while sequence reads having the same barcode are results of PCR duplication from one original molecule. Discover the latest research on molecular barcoding here.

Chronic Fatigue Syndrome

Chronic fatigue syndrome is a disease characterized by unexplained disabling fatigue; the pathology of which is incompletely understood. Discover the latest research on chronic fatigue syndrome here.

Evolution of Pluripotency

Pluripotency refers to the ability of a cell to develop into three primary germ cell layers of the embryo. This feed focuses on the mechanisms that underlie the evolution of pluripotency. Here is the latest research.

Position Effect Variegation

Position Effect Variagation occurs when a gene is inactivated due to its positioning near heterochromatic regions within a chromosome. Discover the latest research on Position Effect Variagation here.

STING Receptor Agonists

Stimulator of IFN genes (STING) are a group of transmembrane proteins that are involved in the induction of type I interferon that is important in the innate immune response. The stimulation of STING has been an active area of research in the treatment of cancer and infectious diseases. Here is the latest research on STING receptor agonists.

Microbicide

Microbicides are products that can be applied to vaginal or rectal mucosal surfaces with the goal of preventing, or at least significantly reducing, the transmission of sexually transmitted infections. Here is the latest research on microbicides.