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Investigation of DNA Repair Pathway Activity

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Dictyostelium discoideum Protocols

Part of the book series: Methods in Molecular Biology ((MIMB,volume 983))

Abstract

DNA is constantly being damaged from endogenous and exogenous sources and efficient repair of different types of DNA lesions is essential for the survival of the organism. Dictyostelium is highly resistant to DNA damage and its genome sequence has revealed the presence of multiple repair pathways conserved with vertebrates but lost in other genetically tractable invertebrate models. As such, Dictyostelium is a powerful model organism to study selected human DNA repair pathways and may provide insights into the molecular basis of how cells become resistant to DNA damage. Here we describe a range of assays used to study DNA repair in Dictyostelium. Genes required for repair of DNA damage can be identified and analyzed by comparing the ability of control or mutant cells to survive exposure to genotoxic agents that induce different types of DNA lesion. We also describe assays that assess the presence of markers for DNA repair within chromatin either in the form of posttranslational modification of proteins at sites of damage or the recruitment of repair factors to DNA lesions. Finally, we also describe more direct assays to assess repair of DNA double-strand breaks by either homologous recombination or non-homologous end joining.

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Authors and Affiliations

  1. Department of Biochemistry, University of Oxford, Oxford, UK

    Anne-Marie C. Couto, Nicholas D. Lakin & Catherine J. Pears

Authors
  1. Anne-Marie C. Couto
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  2. Nicholas D. Lakin
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  3. Catherine J. Pears
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Corresponding author

Correspondence to Catherine J. Pears .

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Editors and Affiliations

  1. , Centre for Biochemistry, University of Cologne, Joseph-Stelzmann-Str. 52, Köln, 50931, Nordrhein-Westfalen, Germany

    Ludwig Eichinger

  2. , Centre for Biomedical Research, University of Hull, Hull, HU6 7RX, Montserrat

    Francisco Rivero

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Couto, AM.C., Lakin, N.D., Pears, C.J. (2013). Investigation of DNA Repair Pathway Activity. In: Eichinger, L., Rivero, F. (eds) Dictyostelium discoideum Protocols. Methods in Molecular Biology, vol 983. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-302-2_16

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  • DOI: https://doi.org/10.1007/978-1-62703-302-2_16

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  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-62703-301-5

  • Online ISBN: 978-1-62703-302-2

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