Abstract
Gene regulation by transcription factors involves complex protein interaction networks, which include chromatin remodeling and modifying proteins as an integral part. Decoding these protein interactions is crucial for our understanding of chromatin-mediated gene regulation. Here, we describe a method for the immunoprecipitation of in planta nuclear protein complexes followed by mass spectrometry (IP-MS) to identify interactions between transcription factors and chromatin remodelers/modifiers in plants. In addition to a step-by-step bench protocol for immunoprecipitation and subsequent mass spectrometry, we provide guidelines and pointers on necessary controls and data analysis approaches.
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References
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Acknowledgments
We thank Sjef Boeren and Twan America for helpful suggestions regarding the data analysis and Tom Brabbs and Richard Immink for critical reading of the manuscript. S.J. is supported by the European Commission Seventh Framework-People-2012-ITN Project EpiTRAITS, GA-316965 (Epigenetic regulation of economically important plant traits) and M.B. by an NWO-Veni grant.
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Jamge, S., Angenent, G.C., Bemer, M. (2018). Identification of In Planta Protein–Protein Interactions Using IP-MS. In: Bemer, M., Baroux, C. (eds) Plant Chromatin Dynamics. Methods in Molecular Biology, vol 1675. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7318-7_18
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DOI: https://doi.org/10.1007/978-1-4939-7318-7_18
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