Abstract
Histone posttranslational modifications play significant roles in regulating chromatin structure and gene expression. One of the histone modifications, histone citrullination, is catalyzed by an enzyme called peptidylarginine deiminase 4 (PAD4, also called PADI4), which converts both histone arginine (Arg) and mono-methyl arginine residues to citrulline. Recent studies have found that histone citrullination counteracts the effect of histone arginine methylation and functions as a repressive marker to turn off gene expression. Here, we describe assays to study histone citrullination by PAD4 in vitro and in vivo. We also describe approaches to measure histone citrullination levels at gene promoters using chromatin immunoprecipitation assay and analyze the effects of PAD4 inhibitor on cell cycle and apoptosis by flow cytometry. These methods would be useful techniques to study this unique histone modification.
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Acknowledgment
The authors would like to thank the Flow Cytometry Facility at the Pennsylvania State University for its technical help. Research in the Wang laboratory is supported by NIH grant R01 CA136856.
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Li, P., Hu, J., Wang, Y. (2012). Methods for Analyzing Histone Citrullination in Chromatin Structure and Gene Regulation. In: Vancura, A. (eds) Transcriptional Regulation. Methods in Molecular Biology, vol 809. Springer, New York, NY. https://doi.org/10.1007/978-1-61779-376-9_31
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DOI: https://doi.org/10.1007/978-1-61779-376-9_31
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