Quantification of DNA repair protein kinetics after γ-irradiation using number and brightness analysis

Salim Abdisalaam, Milan Poudel, David J. Chen, George Alexandrakis

Research output: Chapter in Book/Report/Conference proceedingConference contribution

Abstract

The kinetics of most proteins involved in DNA damage sensing, signaling and repair following ionizing radiation exposure cannot be quantified by current live cell fluorescence microscopy methods. This is because most of these proteins, with only few notable exceptions, do not attach in large numbers at DNA damage sites to form easily detectable foci in microscopy images. As a result a high fluorescence background from freely moving and immobile fluorescent proteins in the nucleus masks the aggregation of proteins at sparse DNA damage sites. Currently, the kinetics of these repair proteins are studied by laser-induced damage and Fluorescence Recovery After Photobleaching that rely on the detectability of high fluorescence intensity spots of clustered DNA damage. We report on the use of Number and Brightness (N&B) analysis methods as a means to monitor kinetics of DNA repair proteins during sparse DNA damage created by γ-irradiation, which is more relevant to cancer treatment than laser-induced clustered damage. We use two key double strand break repair proteins, namely Ku 70/80 and the DNA-dependent protein kinase catalytic subunit (DNA-PKCS), as specific examples to showcase the feasibility of the proposed methods to quantify dose-dependent kinetics for DNA repair proteins after exposure to γ-rays.

Original languageEnglish (US)
Title of host publicationProgress in Biomedical Optics and Imaging - Proceedings of SPIE
Volume7905
DOIs
StatePublished - 2011
EventSingle Molecule Spectroscopy and Imaging IV - San Francisco, CA, United States
Duration: Jan 22 2011Jan 23 2011

Other

OtherSingle Molecule Spectroscopy and Imaging IV
CountryUnited States
CitySan Francisco, CA
Period1/22/111/23/11

Fingerprint

DNA Repair
Luminance
brightness
DNA
Repair
deoxyribonucleic acid
Irradiation
proteins
Proteins
DNA Damage
Kinetics
irradiation
kinetics
damage
Laser damage
Fluorescence
fluorescence
Lasers
DNA-Activated Protein Kinase
Fluorescence Recovery After Photobleaching

Keywords

  • DNA repair
  • double strand break
  • Fluorescence Correlation Spectroscopy
  • Number and Brightness analysis

ASJC Scopus subject areas

  • Atomic and Molecular Physics, and Optics
  • Electronic, Optical and Magnetic Materials
  • Biomaterials
  • Radiology Nuclear Medicine and imaging

Cite this

Abdisalaam, S., Poudel, M., Chen, D. J., & Alexandrakis, G. (2011). Quantification of DNA repair protein kinetics after γ-irradiation using number and brightness analysis. In Progress in Biomedical Optics and Imaging - Proceedings of SPIE (Vol. 7905). [790517] https://doi.org/10.1117/12.873853

Quantification of DNA repair protein kinetics after γ-irradiation using number and brightness analysis. / Abdisalaam, Salim; Poudel, Milan; Chen, David J.; Alexandrakis, George.

Progress in Biomedical Optics and Imaging - Proceedings of SPIE. Vol. 7905 2011. 790517.

Research output: Chapter in Book/Report/Conference proceedingConference contribution

Abdisalaam, S, Poudel, M, Chen, DJ & Alexandrakis, G 2011, Quantification of DNA repair protein kinetics after γ-irradiation using number and brightness analysis. in Progress in Biomedical Optics and Imaging - Proceedings of SPIE. vol. 7905, 790517, Single Molecule Spectroscopy and Imaging IV, San Francisco, CA, United States, 1/22/11. https://doi.org/10.1117/12.873853
Abdisalaam S, Poudel M, Chen DJ, Alexandrakis G. Quantification of DNA repair protein kinetics after γ-irradiation using number and brightness analysis. In Progress in Biomedical Optics and Imaging - Proceedings of SPIE. Vol. 7905. 2011. 790517 https://doi.org/10.1117/12.873853
Abdisalaam, Salim ; Poudel, Milan ; Chen, David J. ; Alexandrakis, George. / Quantification of DNA repair protein kinetics after γ-irradiation using number and brightness analysis. Progress in Biomedical Optics and Imaging - Proceedings of SPIE. Vol. 7905 2011.
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