Cell Signaling Technology

Product Pathways - Chromatin Regulation / Epigenetics

Phospho-HDAC4 (Ser246)/HDAC5 (Ser259)/HDAC7 (Ser155) (D27B5) Rabbit mAb #3443

Applications Reactivity Sensitivity MW (kDa) Isotype
W IP H M Endogenous 140, 124, 120 Rabbit IgG

Applications Key:  W=Western Blotting  IP=Immunoprecipitation
Reactivity Key:  H=Human  M=Mouse
Species cross-reactivity is determined by western blot. Species enclosed in parentheses are predicted to react based on 100% sequence homology.

Protocols

Specificity / Sensitivity

Phospho-HDAC4 (Ser246)/HDAC5 (Ser259)/HDAC7 (Ser155) (D27B5) Rabbit mAb detects endogenous levels of HDAC4, HDAC5 and HDAC7 proteins only when phosphorylated on Ser246, Ser259 and Ser155, respectively.

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic phosphopeptide corresponding to Ser155 of human HDAC7 protein.

Western Blotting

Western Blotting

Western blot analysis of extracts from DO11.10 thymocyte hybridoma cells, either untreated or treated for 1 h with TPA (0.2 uM) and ionomycin (0.33 uM), using Phospho-HDAC4 (Ser246)/HDAC5 (Ser259)/HDAC7 (Ser155) (D27B5) Rabbit mAb. Phospho-specificity of the antibody was determined by treating cell extracts with lambda phosphatase. Total HDAC proteins were detected using Histone Deacetylase 4 (HDAC4) Antibody #2072, Histone Deacetylase 5 (HDAC5) Antibody #2082 and Histone Deacetylase 7 (HDAC7) Antibody #2882.

Background

Acetylation of the histone tail causes chromatin to adopt an "open" conformation, allowing increased accessibility of transcription factors to DNA. The identification of histone acetyltransferases (HATs) and their large multiprotein complexes has yielded important insights into how these enzymes regulate transcription (1,2). HAT complexes interact with sequence-specific activator proteins to target specific genes. In addition to histones, HATs can acetylate nonhistone proteins, suggesting multiple roles for these enzymes (3). In contrast, histone deacetylation promotes a "closed" chromatin conformation and typically leads to repression of gene activity (4). Mammalian histone deacetylases can be divided into three classes on the basis of their similarity to various yeast deacetylases (5). Class I proteins (HDACs 1, 2, 3, and 8) are related to the yeast Rpd3-like proteins, those in class II (HDACs 4, 5, 6, 7, 9, and 10) are related to yeast Hda1-like proteins, and class III proteins are related to the yeast protein Sir2. Inhibitors of HDAC activity are now being explored as potential therapeutic cancer agents (6,7).

Histone deacetylases (HDACs) interact with an increasing number of transcription factors, including myocyte enhancer factor 2 (MEF2), to negatively regulate gene expression. HDACs are regulated in part by shuttling between the nucleus and cytoplasm, where export to the cytoplasm facilitates gene activation by removing HDACs from their target genes (8,9). The cytoplasmic export is facilitated by 14-3-3 proteins, which bind to specific phospho-serine residues on the HDAC proteins (8,9). These phospho-serine 14-3-3 binding modules are highly conserved between HDAC proteins, allowing for their collective regulation in response to specific cell stimuli. For example, the highly conserved HDAC 4 Ser246, HDAC 5 Ser259 and HDAC 7 Ser155 residues are all phosphorylated by CAMK and PKD kinases in response to multiple cell stimuli, including VEGF-induced angiogenesis in endothelial cells, B cell and T cell activation, and differentiation of myoblasts into muscle fiber (10-14).

  1. Marmorstein, R. (2001) Cell Mol Life Sci 58, 693-703.
  2. Gregory, P.D. et al. (2001) Exp Cell Res 265, 195-202.
  3. Liu, Y. et al. (2000) Mol Cell Biol 20, 5540-53.
  4. Cress, W.D. and Seto, E. (2000) J Cell Physiol 184, 1-16.
  5. Gray, S.G. and Ekström, T.J. (2001) Exp Cell Res 262, 75-83.
  6. Thiagalingam, S. et al. (2003) Ann. N.Y. Acad. Sci. 983, 84-100.
  7. Vigushin, D.M. and Coombes, R.C. (2004) Curr. Cancer Drug Targets 4, 205-218.
  8. Grozinger, C.M. and Schreiber, S.L. (2000) Proc Natl Acad Sci U S A 97, 7835-40.
  9. Wang, A.H. et al. (2000) Mol Cell Biol 20, 6904-12.
  10. Ha, C.H. et al. (2008) J Biol Chem 283, 14590-9.
  11. Wang, S. et al. (2008) Proc Natl Acad Sci U S A 105, 7738-43.
  12. Matthews, S.A. et al. (2006) Mol Cell Biol 26, 1569-77.
  13. Parra, M. et al. (2005) J Biol Chem 280, 13762-70.
  14. McKinsey, T.A. et al. (2000) Nature 408, 106-11.

Application References

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This product is intended for research purposes only. The product is not intended to be used for therapeutic or diagnostic purposes in humans or animals.

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