Visualization of DNA Double-Strand Break Repair at the Single-Molecule Level

Conference · Thu Mar 27 04:00:00 EST 2003

OSTI ID:826401

Dynan, William S; Li, Shuyi; Mernaugh, Raymond; Wragg, Stephanie; Takeda, Yoshihiko

Exposure to low doses of ionizing radiation is universal. The signature injury from ionizing radiation exposure is induction of DNA double-strand breaks (DSBs). The first line of defense against DSBs is direct ligation of broken DNA ends via the nonhomologous end-joining pathway. Because even a relatively high environmental exposure induces only a few DSBs per cell, our current understanding of the response to this exposure is limited by the ability to measure DSB repair events reliably in situ at a single-molecule level. To address this need, we have taken advantage of biological amplification, measuring relocalization of proteins and detection of protein phosphorylation as a surrogate for detection of broken ends themselves. We describe the use of specific antibodies to investigate the kinetics and mechanism of repair of very small numbers of DSBs in human cells by the nonhomologous end-joining pathway.

• Research Organization: Medical College of Georgia, Augusta, GA; Vanderbilt University, Nashville, TN (US)
• Sponsoring Organization: USDOE Office of Science (SC) (US)
• OSTI ID: 826401
• Country of Publication: United States
• Language: English

Related Subjects

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CHEMISTRY
54 ENVIRONMENTAL SCIENCES
59 BASIC BIOLOGICAL SCIENCES
60 APPLIED LIFE SCIENCES
AMPLIFICATION
ANIMAL CELLS
ANTIBODIES
DETECTION
DNA
ENVIRONMENTAL EXPOSURE
INDUCTION
IONIZING RADIATIONS
KINETICS
PHOSPHORYLATION
PROTEINS
REPAIR