Biomaterial Database

Analyzing Heterochromatic DNA Double Strand Break (DSB) Repair in Response to Ionizing Radiation. 2017

Karolin Klement, and Aaron A Goodarzi

Robson DNA Science Centre, Arnie Charbonneau Cancer Institute, Department of Biochemistry and Molecular Biology and Department of Oncology, Cumming School of Medicine, University of Calgary, Calgary, AB, Canada, T2N 4N1.

DNA damaging agents such as ionizing irradiation induce lesions in the DNA such as double strand breaks (DSBs). Depending on cell type, 10-25% of these DSBs are induced in heterochromatin. Heterochromatic DSBs are resolved with slow kinetics (compared to DSBs in euchromatin) and require ATM activity for repair. Investigating the underlying causes of the slow component of DSB repair and the role of individual response factors in this process provides insight into DSB response pathways and will further the understanding of diseases where such pathways are dysfunctional due to mutation. Here, we describe a method to detect DSB repair foci in the heterochromatin of human cells. We provide a detailed protocol for cell culture preparation, immunofluorescence microscopy, and a computer-assisted approach to analyze overlap between DSB foci and heterochromatin.

UI	MeSH Term	Description	Entries
D011839	Radiation, Ionizing	ELECTROMAGNETIC RADIATION or particle radiation (high energy ELEMENTARY PARTICLES) capable of directly or indirectly producing IONS in its passage through matter. The wavelengths of ionizing electromagnetic radiation are equal to or smaller than those of short (far) ultraviolet radiation and include gamma and X-rays.	Ionizing Radiation,Ionizing Radiations,Radiations, Ionizing
D004260	DNA Repair	The removal of DNA LESIONS and/or restoration of intact DNA strands without BASE PAIR MISMATCHES, intrastrand or interstrand crosslinks, or discontinuities in the DNA sugar-phosphate backbones.	DNA Damage Response
D006801	Humans	Members of the species Homo sapiens.	Homo sapiens,Man (Taxonomy),Human,Man, Modern,Modern Man
D053903	DNA Breaks, Double-Stranded	Interruptions in the sugar-phosphate backbone of DNA, across both strands adjacently.	Double-Stranded DNA Breaks,Double-Strand DNA Breaks,Double-Stranded DNA Break,Break, Double-Strand DNA,Break, Double-Stranded DNA,Breaks, Double-Strand DNA,Breaks, Double-Stranded DNA,DNA Break, Double-Strand,DNA Break, Double-Stranded,DNA Breaks, Double Stranded,DNA Breaks, Double-Strand,Double Strand DNA Breaks,Double Stranded DNA Break,Double Stranded DNA Breaks,Double-Strand DNA Break
D064007	Ataxia Telangiectasia Mutated Proteins	A group of PROTEIN SERINE-THREONINE KINASES which activate critical signaling cascades in double strand breaks, APOPTOSIS, and GENOTOXIC STRESS such as ionizing ultraviolet A light, thereby acting as a DNA damage sensor. These proteins play a role in a wide range of signaling mechanisms in cell cycle control.	A-T Protein,AT Mutated Protein,ATM Protein,A T Protein,Mutated Protein, AT,Protein, A-T,Protein, AT Mutated