Abstract
Telomere repeats at chromosomal ends are essential for genome stability and sustained cellular proliferation but are susceptible to DNA damage. Repair of damage at telomeres is influenced by numerous factors including telomeric binding proteins, sequence and structure. Ultraviolet (UV) light irradiation induces DNA photoproducts at telomeres that can interfere with telomere maintenance. Here we describe a highly sensitive method for quantifying the formation and removal of UV photoproducts in telomeres isolated from UV irradiated cultured human cells. Damage is detected by immunospot blotting of telomeres with highly specific antibodies against UV photoproducts. This method is adaptable for measuring other types of DNA damage at telomeres as well.
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Acknowledgments
The authors thank Dr. Woodring Wright (University of Texas Southwestern Medical Center) for helpful discussion regarding the telomere isolation protocol and acknowledge Dr. Dhvani Parikh for her contributions toward developing and optimizing the method for UV photoproduct detection at telomeres. This work was supported by an NIH grant (R01ES022944, ES28242, ES025606 to P.L.O, R21ES027641, R01GM107559, R01GM123246 to H.W.), P30 ES025128 (through a pilot project grant to H.W. by CHHE at NCSU), and NIH grant (K99ES027028 to E.F.).
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Fouquerel, E., Barnes, R.P., Wang, H., Opresko, P.L. (2019). Measuring UV Photoproduct Repair in Isolated Telomeres and Bulk Genomic DNA. In: Balakrishnan, L., Stewart, J. (eds) DNA Repair. Methods in Molecular Biology, vol 1999. Humana, New York, NY. https://doi.org/10.1007/978-1-4939-9500-4_20
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DOI: https://doi.org/10.1007/978-1-4939-9500-4_20
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