ArticleNucleic acids research2014
Scanning fluorescence correlation spectroscopy techniques to quantify the kinetics of DNA double strand break repair proteins after γ-irradiation and bleomycin treatment.
Article in Nucleic acids research, 2014. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 10 papers.
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Who cites it
10 citing papers in PubMed.
- Heavy metal ions exchange driven protein phosphorylation cascade functions in genomic instability in spermatocytes and male infertility.Nucleic acids research · 2023Article
- A computational approach to quantifying miscounting of radiation-induced double-strand break immunofluorescent foci.Communications biology · 2022Article
- New Methodologies to Study DNA Repair Processes in Space and Time Within Living Cells.Frontiers in cell and developmental biology · 2021Review
- The nuclear structural protein NuMA is a negative regulator of 53BP1 in DNA double-strand break repair.Nucleic acids research · 2019Article
- Using enhanced number and brightness to measure protein oligomerization dynamics in live cells.Nature protocols · 2019Article
- Differentiated embryo chondrocyte plays a crucial role in DNA damage response via transcriptional regulation under hypoxic conditions.PloS one · 2018Article
- Engineering new balancer chromosomes in C. elegans via CRISPR/Cas9.Scientific reports · 2016Article
- Raster image cross-correlation analysis for spatiotemporal visualization of intracellular degradation activities against exogenous DNAs.Scientific reports · 2015Article
- Organization and dynamics of the nonhomologous end-joining machinery during DNA double-strand break repair.Proceedings of the National Academy of Sciences of the United States of America · 2015Article
- Cytoskeletal regulation of CD44 membrane organization and interactions with E-selectin.The Journal of biological chemistry · 2014Article
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Authors and funding
4 authors.
Funding
Abstract
A common feature of DNA repair proteins is their mobilization in response to DNA damage. The ability to visualizing and quantifying the kinetics of proteins localizing/dissociating from DNA double strand breaks (DSBs) via immunofluorescence or live cell fluorescence microscopy have been powerful tools in allowing insight into the DNA damage response, but these tools have some limitations. For example, a number of well-established DSB repair factors, in particular those required for non-homologous end joining (NHEJ), do not form discrete foci in response to DSBs induced by ionizing radiation (IR) or radiomimetic drugs, including bleomycin, in living cells. In this report, we show that time-dependent kinetics of the NHEJ factors Ku80 and DNA-dependent protein kinase catalytic subunits (DNA-PKcs) in response to IR and bleomycin can be quantified by Number and Brightness analysis and Raster-scan Image Correlation Spectroscopy. Fluorescent-tagged Ku80 and DNA-PKcs quickly mobilized in response to IR and bleomycin treatments consistent with prior reports using laser-generated DSBs. The response was linearly dependent on IR dose, and blocking NHEJ enhanced immobilization of both Ku80 and DNA-PKcs after DNA damage. These findings support the idea of using Number and Brightness and Raster-scan Image Correlation Spectroscopy as methods to monitor kinetics of DSB repair proteins in living cells under conditions mimicking radiation and chemotherapy treatments.
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