Abstract
DNA helicases utilize the energy of nucleotide hydrolysis to unwind the two annealed strands of the DNA helix and are involved in many aspects of DNA metabolism such as replication, recombination, and repair. Chromatin immunoprecipitation (ChIP) has been instrumental in determining the genomic targets of many DNA helicases and DNA helicase-containing complexes including the minichromosome maintenance (Mcm) proteins 2–7, the RecQ helicase Sgs1 as well as the Rvb1 and Rvb2 helicase-containing INO80 and SWR1 chromatin remodeling complexes. Here we describe a ChIP method that has been successfully used to map these proteins at chromosomal double-strand breaks and replication forks in the model organism Saccharomyces cerevisiae.
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Acknowledgments
The authors would like to thank Professor Susan Gasser and members of the Gasser Laboratory for support during the development of these protocols. JC is an Alberta Heritage Foundation for Medical Research Scholar and work in the JC laboratory is funded by grants from the Canadian Institutes for Health Research # MOP-82736 and the Alberta Cancer Board # 23575. HvA is a Human Frontiers Science Program long-term fellow and work in HvA’s group is funded by a VIDI grant from the Netherlands Organisation for Scientific Research (NWO).
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Cobb, J., van Attikum, H. (2009). Mapping Genomic Targets of DNA Helicases by Chromatin Immunoprecipitation in Saccharomyces cerevisiae . In: Abdelhaleem, M. (eds) Helicases. Methods in Molecular Biology, vol 587. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-60327-355-8_8
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DOI: https://doi.org/10.1007/978-1-60327-355-8_8
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