Abstract
One of the most important roles of Sirtuins is to ensure the maintenance of genome integrity under stress conditions. In this chapter, we provide a methodology to study this role of Sirtuins at two different levels: detection of genomic instability (with the Neutral Comet Assay) and study of Sirtuin dynamics in chromatin under stress conditions (by isolating insoluble chromatin fractions).
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References
Lombard DB, Chua KF, Mostoslavsky R, Gostissa M, Franco S, Alt FW (2005) DNA repair, genome stability, and aging. Cell 120:497–512
Guarente L (1999) Diverse and dynamic functions of the Sir silencing complex. Nat Genet 23:281–285
Bosch-Presegué L, Vaquero A (2011) A dual role of sirtuins in cáncer. Genes Cancer 2: 648–662
Imai S, Armstrong CM, Kaeberlein M, Guarente L (2000) Transcriptional silencing and longevity protein Sir2 is an NAD-dependent histone deacetylase. Nature 403: 795–800
Landry J, Sutton A, Tafrov ST et al (2000) The silencing protein SIR2 and its homologs are NADdependent protein deacetylases. Proc Natl Acad Sci U S A 97:5807–5811
Tanny JC, Dowd GJ, Huang J, Hilz H, Moazed D (1999) An enzymatic activity in the yeast Sir2 protein that is essential for gene silencing. Cell 99:735–745
Wang RH, Sengupta K, Li C et al (2008) Impaired DNA damage response, genome instability, and tumorigenesis in SIRT1 mutant mice. Cancer Cell 14:312–323
Mostoslavsky R, Chua KF, Lombard DB et al (2006) Genomic instability and aging-like phenotype in the absence of mammalian SIRT6. Cell 124:315–329
Vaquero A (2009) The conserved role of sirtuins in chromatin regulation. Int J Dev Biol 53:303–322
Webster BR, Lu Z, Sack MN, Scott I (2012) The role of sirtuins in modulating redox stressors. Free Radic Biol Med 52(2):281–290
Ostling O, Johanson KJ (1984) Microelectrophoretic study of radiation-induced DNA damages in individual mammalian cells. Biochem Biophys Res Commun 123:291–298
Hajji N, Wallenborg K, Vlachos P, Füllgrabe J, Hermanson O, Joseph B (2010) Zpposing effects of hMOF and SIRT1 on H4K16 acetylation and the sensitivity to the topoisomerase II inhibitor etoposide. Oncogene 29:2192–2204
McCord RA, Michishita E, Hong T, Berber E, Boxer LD, Kusumoto R, Guan S, Shi X, Gozani O, Burlingame AL, Bohr VA, Chua KF (2009) SIRT6 stabilizes DNA-dependent protein kinase at chromatin for DNA double-strand break repair. Aging 1(1):109–121
Holden P, Horton WA (2009) Crude subcellular fractionation of cultured mammalian cell lines. BMC Res Notes 2:243
Dignam JD, Martin PL, Shastry BS, Roeder RG (1983) Eukaryotic gene transcription with purified components. Methods Enzymol 101:582–598
Olive PL, Banáth JP (2006) The comet assay: a method to measure DNA damage in individual cells. Nat Protoc 1:23–29
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Martínez-Redondo, P., Vaquero, A. (2013). Methods to Study the Role of Sirtuins in Genome Stability. In: Hirschey, M. (eds) Sirtuins. Methods in Molecular Biology, vol 1077. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-637-5_18
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DOI: https://doi.org/10.1007/978-1-62703-637-5_18
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Publisher Name: Humana Press, Totowa, NJ
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