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).
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
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
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2013 Springer Science+Business Media, LLC
About this protocol
Cite this protocol
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
Download citation
DOI: https://doi.org/10.1007/978-1-62703-637-5_18
Published:
Publisher Name: Humana Press, Totowa, NJ
Print ISBN: 978-1-62703-636-8
Online ISBN: 978-1-62703-637-5
eBook Packages: Springer Protocols