A Sensitive and Flexible Assay for Determining Histone Deacetylase 1 (HDAC1) Activity

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

Abstract

Histones acetylation and deacetylation constitute part of the so-called “histone code” and work in concert with other posttranslational modifications to determine the activity of genes. Deacetylation of histone is carried out by a class of enzymes, known as histone deacetylases (HDACs). The action of HDAC is countered by histone acetyltransferases. Although histone is the best characterized substrate of HDACs, increasing evidence also indicates that non-histone proteins are equally important subtract of HDACs. Since HDACs play an important role in normal physiological and pathophysiological conditions, a sensitive and flexible deacetylation assay that can reliably detect HDAC activity and identify potential novel targets of HDACs is critical.

Key words

HDAC1 Deacetylation Immunoaffinity purification Core histone Posttranslational modification Colorimetric assay 

Notes

Acknowledgements

Work in the authors’ laboratory is supported by grants CA187857 and CA188471 from National Cancer Institute, and McCormick Genomic and Proteomic Center at the George Washington University.

References

  1. 1.
    Strahl BD, Allis CD (2000) The language of covalent histone modifications. Nature 403:41–45CrossRefPubMedGoogle Scholar
  2. 2.
    Leipe DD, Landsman D (1997) Histone deacetylases, acetoin utilization proteins and acetylpolyamine amidohydrolases are members of an ancient protein superfamily. Nucleic Acids Res 25:3693–3697CrossRefPubMedPubMedCentralGoogle Scholar
  3. 3.
    Gregoretti IV, Lee YM, Goodson HV (2004) Molecular evolution of the histone deacetylase family: functional implications of phylogenetic analysis. J Mol Biol 338:17–31CrossRefPubMedGoogle Scholar
  4. 4.
    Taunton J, Hassig CA, Schreiber SL (1996) A mammalian histone deacetylase related to the yeast transcriptional regulator Rpd3p. Science 272:408–411CrossRefPubMedGoogle Scholar
  5. 5.
    de Ruijter AJ, van Gennip AH, Caron HN, Kemp S, van Kuilenburg AB (2003) Histone deacetylases (HDACs): characterization of the classical HDAC family. Biochem J 370:737–749CrossRefPubMedPubMedCentralGoogle Scholar
  6. 6.
    Takami Y, Nakayama T (2000) N-terminal region, C-terminal region, nuclear export signal, and deacetylation activity of histone deacetylase-3 are essential for the viability of the DT40 chicken B cell line. J Biol Chem 275:16191–16201CrossRefPubMedGoogle Scholar
  7. 7.
    Yang WM, Tsai SC, Wen YD, Fejer G, Seto E (2002) Functional domains of histone deacetylase-3. J Biol Chem 277:9447–9454CrossRefPubMedGoogle Scholar
  8. 8.
    Witt O, Deubzer HE, Milde T, Oehme I (2009) HDAC family: what are the cancer relevant targets? Cancer Lett 277:8–21CrossRefPubMedGoogle Scholar
  9. 9.
    Haigis MC, Guarente LP (2006) Mammalian sirtuins—emerging roles in physiology, aging, and calorie restriction. Genes Dev 20:2913–2921CrossRefPubMedGoogle Scholar
  10. 10.
    Gao L, Cueto MA, Asselbergs F, Atadja P (2002) Cloning and functional characterization of HDAC11, a novel member of the human histone deacetylase family. J Biol Chem 277:25748–25755CrossRefPubMedGoogle Scholar
  11. 11.
    Senese S, Zaragoza K, Minardi S, Muradore I, Ronzoni S, Passafaro A, Bernard L, Draetta GF, Alcalay M, Seiser C, Chiocca S (2007) Role for histone deacetylase 1 in human tumor cell proliferation. Mol Cell Biol 27:4784–4795CrossRefPubMedPubMedCentralGoogle Scholar
  12. 12.
    Zupkovitz G, Grausenburger R, Brunmeir R, Senese S, Tischler J, Jurkin J, Rembold M, Meunier D, Egger G, Lagger S, Chiocca S, Propst F, Weitzer G, Seiser C (2010) The cyclin-dependent kinase inhibitor p21 is a crucial target for histone deacetylase 1 as a regulator of cellular proliferation. Mol Cell Biol 30:1171–1181CrossRefPubMedGoogle Scholar
  13. 13.
    Haberland M, Montgomery RL, Olson EN (2009) The many roles of histone deacetylases in development and physiology: implications for disease and therapy. Nat Rev Genet 10:32–42CrossRefPubMedPubMedCentralGoogle Scholar
  14. 14.
    Lagger G, O'Carroll D, Rembold M, Khier H, Tischler J, Weitzer G, Schuettengruber B, Hauser C, Brunmeir R, Jenuwein T, Seiser C (2002) Essential function of histone deacetylase 1 in proliferation control and CDK inhibitor repression. EMBO J 21:2672–2681CrossRefPubMedPubMedCentralGoogle Scholar
  15. 15.
    Karwowska-Desaulniers P, Ketko A, Kamath N, Pflum MK (2007) Histone deacetylase 1 phosphorylation at S421 and S423 is constitutive in vivo, but dispensable in vitro. Biochem Biophys Res Commun 361:349–355CrossRefPubMedGoogle Scholar
  16. 16.
    Pflum MK, Tong JK, Lane WS, Schreiber SL (2001) Histone deacetylase 1 phosphorylation promotes enzymatic activity and complex formation. J Biol Chem 276:47733–47741CrossRefPubMedGoogle Scholar
  17. 17.
    Rush J, Moritz A, Lee KA, Guo A, Goss VL, Spek EJ, Zhang H, Zha XM, Polakiewicz RD, Comb MJ (2005) Immunoaffinity profiling of tyrosine phosphorylation in cancer cells. Nat Biotechnol 23:94–101CrossRefPubMedGoogle Scholar
  18. 18.
    Sun JM, Chen HY, Davie JR (2007) Differential distribution of unmodified and phosphorylated histone deacetylase 2 in chromatin. J Biol Chem 282:33227–33236CrossRefPubMedGoogle Scholar
  19. 19.
    Tsai SC, Seto E (2002) Regulation of histone deacetylase 2 by protein kinase CK2. J Biol Chem 277:31826–31833CrossRefPubMedGoogle Scholar
  20. 20.
    Wu MY, Fu J, Xiao X, Wu J, Wu RC (2014) MiR-34a regulates therapy resistance by targeting HDAC1 and HDAC7 in breast cancer. Cancer Lett 354:311–319CrossRefPubMedGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2016

Authors and Affiliations

  1. 1.Department of Biochemistry and Molecular MedicineThe George Washington UniversityWashingtonUSA
  2. 2.Department of Molecular and Cellular BiologyBaylor College of MedicineHoustonUSA

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