TY - CHAP
T1 - Analyzing cell cycle checkpoints in response to ionizing radiation in mammalian cells
AU - Wang, Bin
PY - 2014
Y1 - 2014
N2 - Exposure of cells to DNA-damaging agents, such as ionizing radiation (IR), results in perturbation of cell cycle progression. IR activates cell cycle checkpoints that arrest the cell cycle at the G1/S, S, and G2/M phases. The DNA damage-signaling network involves a number of important DNA damage response factors that are required for maintaining genome stability and prevention of cancer. These factors are involved in the regulation of cell cycle checkpoints and include ATM, NBS1, BRCA1, Chk2, and p53. Here we describe a series of assays that are often used to analyze cell cycle checkpoints after IR. These assays include a G1/S checkpoint assay that measures 5-bromodeoxyuridine incorporation into DNA, an S-phase checkpoint assay that measures DNA synthesis at a very early time point after IR, and a G2/M checkpoint assay that quantitates histone H3 phosphorylation. This collection of assays allows us to investigate the specific functions of proteins involved in regulating different cell cycle checkpoints in mammalian cells as a response to IR.
AB - Exposure of cells to DNA-damaging agents, such as ionizing radiation (IR), results in perturbation of cell cycle progression. IR activates cell cycle checkpoints that arrest the cell cycle at the G1/S, S, and G2/M phases. The DNA damage-signaling network involves a number of important DNA damage response factors that are required for maintaining genome stability and prevention of cancer. These factors are involved in the regulation of cell cycle checkpoints and include ATM, NBS1, BRCA1, Chk2, and p53. Here we describe a series of assays that are often used to analyze cell cycle checkpoints after IR. These assays include a G1/S checkpoint assay that measures 5-bromodeoxyuridine incorporation into DNA, an S-phase checkpoint assay that measures DNA synthesis at a very early time point after IR, and a G2/M checkpoint assay that quantitates histone H3 phosphorylation. This collection of assays allows us to investigate the specific functions of proteins involved in regulating different cell cycle checkpoints in mammalian cells as a response to IR.
KW - Brdu
KW - Cell cycle checkpoints
KW - DNA synthesis
KW - Flow cytometry
KW - Histone H3 phosphorylation
KW - Ionizing radiation
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U2 - 10.1007/978-1-4939-888-2_15
DO - 10.1007/978-1-4939-888-2_15
M3 - Chapter
AN - SCOPUS:84934440989
SN - 9781493908875
T3 - Methods in Molecular Biology
SP - 313
EP - 320
BT - Cell Cycle Control
PB - Humana Press Inc.
ER -