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Approaches for Studying the Subcellular Localization, Interactions, and Regulation of Histone Deacetylase 5 (HDAC5)

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Histone Deacetylases

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1436))

Abstract

As a member of the class IIa family of histone deacetylases, the histone deacetylase 5 (HDAC5) is known to undergo nuclear–cytoplasmic shuttling and to be a critical transcriptional regulator. Its misregulation has been linked to prominent human diseases, including cardiac diseases and tumorigenesis. In this chapter, we describe several experimental methods that have proven effective for studying the functions and regulatory features of HDAC5. We present methods for assessing the subcellular localization, protein interactions, posttranslational modifications (PTMs), and activity of HDAC5 from the standpoint of investigating either the endogenous protein or tagged protein forms in human cells. Specifically, given that at the heart of HDAC5 regulation lie its dynamic localization, interactions, and PTMs, we present methods for assessing HDAC5 localization in fixed and live cells, for isolating HDAC5-containing protein complexes to identify its interactions and modifications, and for determining how these PTMs map to predicted HDAC5 structural motifs. Lastly, we provide examples of approaches for studying HDAC5 functions with a focus on its regulation during cell-cycle progression. These methods can readily be adapted for the study of other HDACs or non-HDAC-proteins of interest. Individually, these techniques capture temporal and spatial snapshots of HDAC5 functions; yet together, these approaches provide powerful tools for investigating both the regulation and regulatory roles of HDAC5 in different cell contexts relevant to health and disease.

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Acknowledgments

We are grateful to the current and past members of the Cristea Lab for development and refinement of the techniques discussed in this chapter. This work was supported by grants from the National Institutes of Health (DP1 DA026192 and R01 HL127640) to IMC and an NSF Graduate Research Fellowship to AJG.

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  1. Department of Molecular Biology, Princeton University, 210 Lewis Thomas Laboratory, Princeton, NJ, 08544, USA

    Amanda J. Guise & Ileana M. Cristea

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  1. Amanda J. Guise
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Correspondence to Ileana M. Cristea .

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  1. Boston University School of Medicine, Boston, Massachusetts, USA

    Sibaji Sarkar

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Guise, A.J., Cristea, I.M. (2016). Approaches for Studying the Subcellular Localization, Interactions, and Regulation of Histone Deacetylase 5 (HDAC5). In: Sarkar, S. (eds) Histone Deacetylases. Methods in Molecular Biology, vol 1436. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-3667-0_5

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  • DOI: https://doi.org/10.1007/978-1-4939-3667-0_5

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