Chromatin Immunoprecipitation (ChIP) of Histone Modifications from Saccharomyces cerevisiae

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dc.contributor.authorJezek, Meagan
dc.contributor.authorJacques, Alison
dc.contributor.authorJaiswal, Deepika
dc.contributor.authorGreen, Erin M.
dc.date.accessioned2018-02-16T13:39:08Z
dc.date.available2018-02-16T13:39:08Z
dc.date.issued2017
dc.description.abstractHistone post-translational modifications (PTMs), such as acetylation, methylation and phosphorylation, are dynamically regulated by a series of enzymes that add or remove these marks in response to signals received by the cell. These PTMS are key contributors to the regulation of processes such as gene expression control and DNA repair. Chromatin immunoprecipitation (chIP) has been an instrumental approach for dissecting the abundance and localization of many histone PTMs throughout the genome in response to diverse perturbations to the cell. Here, a versatile method for performing chIP of post-translationally modified histones from the budding yeast Saccharomyces cerevisiae (S. cerevisiae) is described. This method relies on crosslinking of proteins and DNA using formaldehyde treatment of yeast cultures, generation of yeast lysates by bead beating, solubilization of chromatin fragments by micrococcal nuclease, and immunoprecipitation of histone-DNA complexes. DNA associated with the histone mark of interest is purified and subjected to quantitative PCR analysis to evaluate its enrichment at multiple loci throughout the genome. Representative experiments probing the localization of the histone marks H3K4me2 and H4K16ac in wildtype and mutant yeast are discussed to demonstrate data analysis and interpretation. This method is suitable for a variety of histone PTMs and can be performed with different mutant strains or in the presence of diverse environmental stresses, making it an excellent tool for investigating changes in chromatin dynamics under different conditions.en_US
dc.description.urihttps://www.jove.com/video/57080en_US
dc.format.extent8 pagesen_US
dc.identifierdoi:10.13016/M2G44HS3V
dc.identifier.citationJezek, M., Jacques, A., Jaiswal, D., Green, E.M. Chromatin Immunoprecipitation (ChIP) of Histone Modifications from Saccharomyces cerevisiae. Journal of Visualized Experienc, e57080, doi:10.3791/57080 (2017).en_US
dc.identifier.uridoi:10.3791/57080
dc.identifier.urihttp://hdl.handle.net/11603/7797
dc.language.isoen_USen_US
dc.publisherJoveen_US
dc.relation.isAvailableAtThe University of Maryland, Baltimore County (UMBC)
dc.relation.ispartofUMBC Biological Sciences Department Collection
dc.relation.ispartofUMBC Faculty Collection
dc.relation.ispartofUMBC Student Collection
dc.rightsThis item may be protected under Title 17 of the U.S. Copyright Law. It is made available by UMBC for non-commercial research and education. For permission to publish or reproduce, please contact the author.
dc.subjectEpigeneticsen_US
dc.subjectchromatinen_US
dc.subjecthistone modificationsen_US
dc.subjectmethylationen_US
dc.subjectacetylationen_US
dc.subjectchromatin immunoprecipitationen_US
dc.subjectyeasten_US
dc.titleChromatin Immunoprecipitation (ChIP) of Histone Modifications from Saccharomyces cerevisiaeen_US
dc.typeCollectionen_US

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