Abstract
Histone acetylation is dynamically and reversibly controlled by histone acetyltransferases and deacetylases during cellular events such as cell division and differentiation. However, the dynamics of histone modifications in living cells are poorly understood because of the lack of experimental tools to monitor them in a real-time fashion. Herein, we introduce Förster/fluorescence resonance energy transfer (FRET)-based indicators to visualize acetylation of histone H4, and describe a protocol for live-cell imaging with high spatiotemporal resolution.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Muller S, Filippakopoulos P, Knapp S (2011) Bromodomains as therapeutic targets. Expert Rev Mol Med 13:e29
Miyawaki A (2011) Development of probes for cellular functions using fluorescent proteins and fluorescence resonance energy transfer. Annu Rev Biochem 80:357–373
Sasaki K, Ito T, Nishino N, Khochbin S, Yoshida M (2009) Real-time imaging of histone H4 hyperacetylation in living cells. Proc Natl Acad Sci U S A 106:16257–16262
Nguyen AW, Daugherty PS (2005) Evolutionary optimization of fluorescent proteins for intracellular FRET. Nat Biotechnol 23:355–360
Nagai T, Yamada S, Tominaga T, Ichikawa M, Miyawaki A (2004) Expanded dynamic range of fluorescent indicators for Ca(2+) by circularly permuted yellow fluorescent proteins. Proc Natl Acad Sci U S A 101:10554–10559
Komatsu N, Aoki K, Yamada M, Yukinaga H, Fujita Y, Kamioka Y et al (2011) Development of an optimized backbone of FRET biosensors for kinases and GTPases. Mol Biol Cell 22:4647–4656
Hassan AH, Awad S, Al-Natour Z, Othman S, Mustafa F, Rizvi TA (2007) Selective recognition of acetylated histones by bromodomains in transcriptional co-activators. Biochem J 402:125–133
Kanno T, Kanno Y, Siegel RM, Jang MK, Lenardo MJ, Ozato K (2004) Selective recognition of acetylated histones by bromodomain proteins visualized in living cells. Mol Cell 13:33–43
Pivot-Pajot C, Caron C, Govin J, Vion A, Rousseaux S, Khochbin S (2003) Acetylation-dependent chromatin reorganization by BRDT, a testis-specific bromodomain-containing protein. Mol Cell Biol 23:5354–5365
Moriniere J, Rousseaux S, Steuerwald U, Soler-Lopez M, Curtet S, Vitte AL et al (2009) Cooperative binding of two acetylation marks on a histone tail by a single bromodomain. Nature 461:664–668
Umehara T, Nakamura Y, Jang MK, Nakano K, Tanaka A, Ozato K et al (2010) Structural basis for acetylated histone H4 recognition by the human BRD2 bromodomain. J Biol Chem 285:7610–7618
Ito T, Umehara T, Sasaki K, Nakamura Y, Nishino N, Terada T et al (2011) Real-time imaging of histone H4K12-specific acetylation determines the modes of action of histone deacetylase and bromodomain inhibitors. Chem Biol 18:495–507
Shimozono S, Miyawaki A (2008) Engineering FRET constructs using CFP and YFP. Methods Cell Biol 85:381–393
Tsien RY, Harootunian AT (1990) Practical design criteria for a dynamic ratio imaging system. Cell Calcium 11:93–109
Acknowledgments
This work was supported by JST, PRESTO.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2014 Springer Science+Business Media, LLC
About this protocol
Cite this protocol
Sasaki, K., Yoshida, M. (2014). Genetically Encoded FRET Indicators for Live-Cell Imaging of Histone Acetylation. In: Zhang, J., Ni, Q., Newman, R. (eds) Fluorescent Protein-Based Biosensors. Methods in Molecular Biology, vol 1071. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-622-1_12
Download citation
DOI: https://doi.org/10.1007/978-1-62703-622-1_12
Published:
Publisher Name: Humana Press, Totowa, NJ
Print ISBN: 978-1-62703-621-4
Online ISBN: 978-1-62703-622-1
eBook Packages: Springer Protocols