Monitoring the DNA Damage Response at Dysfunctional Telomeres
Monitoring the DNA Damage Response at Dysfunctional Telomeres
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DOI：
10.1007/978-1-4939-2963-4_14
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发表时间：
2015-01-01
期刊：
影响因子：
--
通讯作者：
Chang, Sandy
中科院分区：
文献类型：
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作者：
Rai, Rekha;Chang, Sandy
端粒是重复的 DNA 重复序列，覆盖所有真核染色体的末端。它们的正确维护对于基因组稳定性和细胞活力至关重要。端粒功能失调可能是由于端粒 DNA 的自然损耗或由于庇护蛋白成分的去除而产生的。这些未加帽的染色体末端被 DDR 途径识别为 DSB，导致 DNA 损伤传感器在端粒处积累。这些 DDR 蛋白与功能失调的端粒结合形成端粒功能障碍诱导的 DNA 损伤灶 (TIF)。端粒 TIF 的检测提供了量化端粒功能障碍程度和监测下游 DNA 损伤信号通路的机会。在这里，我们描述了一种使用荧光原位杂交 (FISH) 方法检测 TIF 的方法。
Telomeres are repetitive DNA repeats that cap the ends of all eukaryotic chromosomes. Their proper maintenance is essential for genomic stability and cellular viability. Dysfunctional telomeres could arise through natural attrition of telomeric DNA or due to the removal of shelterin components. These uncapped chromosomal ends are recognized as DSBs by the DDR pathway, leading to the accumulation of DNA damage sensors at telomeres. The association of these DDR proteins with dysfunctional telomeres forms telomere dysfunction induced DNA damage foci (TIFs). Detection of TIFs at telomeres provides an opportunity to quantify the extent of telomere dysfunction and monitor downstream DNA damage signaling pathways. Here we describe a method for the detection of TIFs using a fluorescent in situ hybridization (FISH) approach.