Regulation of histone methylation

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In eukaryotes DNA is packaged into chromatin, a nucleoprotein structure consisting of DNA, histones, and non-histone proteins. Chromatin is important for the fidelity of gene expression and its structure can be regulated by the addition of post-translational modifications (PTMs) to histones. One such modification, methylation of histone H3 at lysine 36, is targeted to active genes by the passage of RNA polymerase II, and this PTM is arguably the most recognized hallmark of productive transcription in all eukaryotes. However, despite over a decade worth of research into the mechanisms behind the establishment and function of H3K36 methylation, there is surprisingly little known about how this modification is removed following gene repression. The research proposed in this application will reveal the enzymes and pathways involved in removal of H3K36 methylation during gene repression. We will use S. cerevisiae as a model organism due to the excellent conservation of genes involved in regulation of H3K36 methylation between yeast and other eukaryotes, and the genetic tractability of this model organism. The techniques used include ChIP (chromatin immunoprecipitation), qPCR (quantitative polymerase chain reaction), library preparation for deep sequencing, bioinformatics, protein purification, enzyme kinetics, and yeast genetics, and thus students supported by my NSERC grant will learn a diverse set of skills that are highly sought after in multiple fields of life science research.