Abstract
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.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
Similar content being viewed by others
References
Cosme-Blanco W, Shen MF, Lazar A et al (2007) Telomere dysfunction suppresses spontaneous tumorigenesis in vivo by activating p53-mediated cellular senescence. EMBO Rep 8:497–503
de Lange T (2005) Shelterin: the protein complex that shapes and safeguards human telomeres. Genes Dev 19:2100–2110
Deng Y, Chan SS, Chang S (2008) Telomere dysfunction and tumour suppression: the senescence connection. Nat Rev Cancer 8:450–458
Deng Y, Guo X, Ferguson DO et al (2009) Multiple roles for MRE11 at uncapped telomeres. Nature 460:914–918
Rai R, Zheng H, He H et al (2010) The function of classical and alternative non-homologous end-joining pathways in the fusion of dysfunctional telomeres. EMBO J 29:2598–2610
d'Adda di Fagagna F, Reaper PM, Clay-Farrace L et al (2003) A DNA damage checkpoint response in telomere initiated senescence. Nature 426:194–198
Takai H, Smogorzewska A, de Lange T (2003) DNA damage foci at dysfunctional telomeres. Curr Biol 13:1549–1556
Liu D, Safari A, O’Connor MS et al (2004) PTOP interacts with POT1 and regulates its localization to telomeres. Nat Cell Biol 6:673–680
Guo X, Deng Y, Lin Y et al (2007) Dysfunctional telomeres activate an ATM-ATR-dependent DNA damage response to suppress tumorigenesis. EMBO J 26:4709–4719
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2015 Springer Science+Business Media New York
About this protocol
Cite this protocol
Rai, R., Chang, S. (2015). Monitoring the DNA Damage Response at Dysfunctional Telomeres. In: Shaw, A. (eds) Immunosenescence. Methods in Molecular Biology, vol 1343. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-2963-4_14
Download citation
DOI: https://doi.org/10.1007/978-1-4939-2963-4_14
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-2962-7
Online ISBN: 978-1-4939-2963-4
eBook Packages: Springer Protocols