Direct Interaction of Ca2+/Calmodulin Inhibits Histone Deacetylase 5 Repressor Core Binding to Myocyte Enhancer Factor 2*

Myocyte enhancer factor 2 (MEF2) proteins play a pivotal role in the differentiation of cardiac and skeletal muscle cells. MEF2 factors are regulated by histone deacetylase enzymes such as histone deacetylase 5 (HDAC5). HDAC5 in turn is responsive to Ca2+ signaling mediated by the intracellular calcium sensor calmodulin. Here a combination of proteolytic fragmentation, matrix-assisted laser desorption ionization mass spectrometry, Edman degradation, circular dichroism, gel filtration, and surface plasmon resonance studies is utilized to define and characterize a stable core domain of HDAC5 and to examine its interactions with MEF2a and calmodulin. Results from real time binding experiments provide evidence for direct interaction of Ca2+/calmodulin with HDAC5 inhibiting MEF2a association with this enzyme.

[1]  J. Martín,et al.  Mutational analysis of the DNA binding, dimerization, and transcriptional activation domains of MEF2C , 1996, Molecular and cellular biology.

[2]  D. Bouchez,et al.  A Novel Family of Calmodulin-binding Transcription Activators in Multicellular Organisms* , 2002, The Journal of Biological Chemistry.

[3]  F. Hartl,et al.  Binding of defined regions of a polypeptide to GroEL and its implications for chaperonin-mediated protein folding , 1995, Nature Structural Biology.

[4]  K. Deisseroth,et al.  Translocation of calmodulin to the nucleus supports CREB phosphorylation in hippocampal neurons , 1998, Nature.

[5]  M. Billingsley,et al.  A rapid and sensitive method for detection and quantification of calcineurin and calmodulin-binding proteins using biotinylated calmodulin. , 1985, Proceedings of the National Academy of Sciences of the United States of America.

[6]  Chun Li Zhang,et al.  Class II Histone Deacetylases Act as Signal-Responsive Repressors of Cardiac Hypertrophy , 2002, Cell.

[7]  Wen‐Ming Yang,et al.  Transcriptional repression by YY1 is mediated by interaction with a mammalian homolog of the yeast global regulator RPD3. , 1996, Proceedings of the National Academy of Sciences of the United States of America.

[8]  R. Treisman,et al.  Human SRF-related proteins: DNA-binding properties and potential regulatory targets. , 1991, Genes & development.

[9]  M. Grunstein.  Histone acetylation in chromatin structure and transcription , 1997, Nature.

[10]  N. Bertos,et al.  Class II histone deacetylases: structure, function, and regulation. , 2001, Biochemistry and cell biology = Biochimie et biologie cellulaire.

[11]  L. Guarente,et al.  Sir2 links chromatin silencing, metabolism, and aging. , 2000, Genes & development.

[12]  T. Tsukiyama.  The in vivo functions of ATP-dependent chromatin-remodelling factors , 2002, Nature Reviews Molecular Cell Biology.

[13]  E. Olson,et al.  Identification of a Signal-Responsive Nuclear Export Sequence in Class II Histone Deacetylases , 2001, Molecular and Cellular Biology.

[14]  P. Becker,et al.  Histone acetylation: a switch between repressive and permissive chromatin , 2002, EMBO reports.

[15]  K. Min,et al.  Regulation of lifespan by histone deacetylase , 2002, Ageing Research Reviews.

[16]  M. Rosenberg,et al.  Efficient Expression of Heterologous Genes in Escherichia coli , 1986 .

[17]  S. Schreiber,et al.  A Mammalian Histone Deacetylase Related to the Yeast Transcriptional Regulator Rpd3p , 1996, Science.

[18]  T. Richmond,et al.  Novel dimerization fold of RAP30/RAP74 in human TFIIF at 1.7 A resolution. , 2000, Journal of molecular biology.

[19]  Richard A. Rifkind,et al.  Cloning and characterization of a histone deacetylase, HDAC9 , 2001, Proceedings of the National Academy of Sciences of the United States of America.

[20]  Magnus Holm,et al.  Calcium/calmodulin inhibition of basic-helix-loop-helix transcription factor domains , 1994, Nature.

[21]  A. R. Penheiter,et al.  Tryptophan 1093 Is Largely Responsible for the Slow Off Rate of Calmodulin from Plasma Membrane Ca2+ Pump 4b* , 2002, The Journal of Biological Chemistry.

[22]  T. Richmond,et al.  Crystal structure of MEF2A core bound to DNA at 1.5 A resolution. , 2000, Journal of molecular biology.

[23]  T. Richmond,et al.  Crystal structure of the nucleosome core particle at 2.8 Å resolution , 1997, Nature.

[24]  T. Richmond,et al.  Solvent mediated interactions in the structure of the nucleosome core particle at 1.9 a resolution. , 2002, Journal of molecular biology.

[25]  J. Adelman,et al.  Structure of the gating domain of a Ca2+-activated K+ channel complexed with Ca2+/calmodulin , 2001, Nature.

[26]  C. Allis,et al.  Translating the Histone Code , 2001, Science.

[27]  A. Sharrocks,et al.  The MADS-box family of transcription factors. , 1995, European journal of biochemistry.

[28]  Ya-Li Yao,et al.  Isolation and Characterization of cDNAs Corresponding to an Additional Member of the Human Histone Deacetylase Gene Family* , 1997, The Journal of Biological Chemistry.

[29]  J. Widom.  Structure, dynamics, and function of chromatin in vitro. , 1998, Annual review of biophysics and biomolecular structure.

[30]  Masaya Orita,et al.  A novel target recognition revealed by calmodulin in complex with Ca2+-calmodulin-dependent kinase kinase , 1999, Nature Structural Biology.

[31]  S. Schreiber,et al.  Regulation of histone deacetylase 4 and 5 and transcriptional activity by 14-3-3-dependent cellular localization. , 2000, Proceedings of the National Academy of Sciences of the United States of America.

[32]  Barry I. Posner,et al.  Regulation of Histone Deacetylase 4 by Binding of 14-3-3 Proteins , 2000, Molecular and Cellular Biology.

[33]  T. Ekström,et al.  The human histone deacetylase family. , 2001, Experimental cell research.

[34]  G. Lukács,et al.  Limited proteolysis as a probe for arrested conformational maturation of ΔF508 CFTR , 1998, Nature Structural Biology.

[35]  A. Verdel,et al.  Identification of a New Family of Higher Eukaryotic Histone Deacetylases , 1999, The Journal of Biological Chemistry.

[36]  R. Passier,et al.  CaM kinase signaling induces cardiac hypertrophy and activates the MEF2 transcription factor in vivo. , 2000, The Journal of clinical investigation.

[37]  E. Olson,et al.  A new myocyte-specific enhancer-binding factor that recognizes a conserved element associated with multiple muscle-specific genes. , 1989, Molecular and cellular biology.

[38]  R. Evans,et al.  Isolation and Characterization of Mammalian HDAC10, a Novel Histone Deacetylase* , 2002, The Journal of Biological Chemistry.

[39]  A. Rhoads,et al.  Sequence motifs for calmodulin recognition , 1997, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[40]  Ricky W. Johnstone,et al.  Histone-deacetylase inhibitors: novel drugs for the treatment of cancer , 2002, Nature Reviews Drug Discovery.

[41]  A. Verdel,et al.  mHDA1/HDAC5 Histone Deacetylase Interacts with and Represses MEF2A Transcriptional Activity* , 2000, The Journal of Biological Chemistry.

[42]  G M Clore,et al.  Solution structure of the MEF2A–DNA complex: structural basis for the modulation of DNA bending and specificity by MADS‐box transcription factors , 2000, The EMBO journal.

[43]  E. Miska,et al.  HDAC4 deacetylase associates with and represses the MEF2 transcription factor , 1999, The EMBO journal.

[44]  I. Dawid,et al.  Isolation and characterization of calmodulin genes from Xenopus laevis , 1984, Molecular and cellular biology.

[45]  S. Schreiber,et al.  Three proteins define a class of human histone deacetylases related to yeast Hda1p. , 1999, Proceedings of the National Academy of Sciences of the United States of America.

[46]  E. Olson,et al.  MEF2: a calcium-dependent regulator of cell division, differentiation and death. , 2002, Trends in biochemical sciences.

[47]  Katherine Luby-Phelps,et al.  Ca2+-regulated Dynamic Compartmentalization of Calmodulin in Living Smooth Muscle Cells (*) , 1995, The Journal of Biological Chemistry.

[48]  E. Miska,et al.  MEF‐2 function is modified by a novel co‐repressor, MITR , 1999, The EMBO journal.

[49]  H. Vogel,et al.  Metal ion and drug binding to proteolytic fragments of calmodulin: proteolytic, cadmium-113, and proton nuclear magnetic resonance studies. , 1984, Biochemistry.

[50]  C. Grozinger,et al.  Calcium Regulates Transcriptional Repression of Myocyte Enhancer Factor 2 by Histone Deacetylase 4* , 2000, The Journal of Biological Chemistry.

[51]  P. Wade.  Transcriptional control at regulatory checkpoints by histone deacetylases: molecular connections between cancer and chromatin. , 2001, Human molecular genetics.

[52]  M E Greenberg,et al.  Neuronal activity-dependent cell survival mediated by transcription factor MEF2. , 1999, Science.

[53]  V. Kiermer,et al.  The emerging role of class II histone deacetylases. , 2001, Biochemistry and cell biology = Biochimie et biologie cellulaire.

[54]  T. Grundström,et al.  A novel type of calmodulin interaction in the inhibition of basic helix-loop-helix transcription factors. , 2000, Biochemistry.

[55]  N. Raikhel,et al.  Protein import into the nucleus: an integrated view. , 1995, Annual review of cell and developmental biology.

[56]  P. V. von Hippel,et al.  Calculation of protein extinction coefficients from amino acid sequence data. , 1989, Analytical biochemistry.

[57]  P. Atadja,et al.  Isolation and Characterization of a Novel Class II Histone Deacetylase, HDAC10* , 2002, The Journal of Biological Chemistry.

[58]  H. Youn,et al.  Apoptosis of T cells mediated by Ca2+-induced release of the transcription factor MEF2. , 1999, Science.

[59]  E. Olson,et al.  Signal-dependent activation of the MEF2 transcription factor by dissociation from histone deacetylases. , 2000, Proceedings of the National Academy of Sciences of the United States of America.

[60]  C. Allis,et al.  The language of covalent histone modifications , 2000, Nature.

[61]  H. Heng,et al.  HDAC4, a Human Histone Deacetylase Related to Yeast HDA1, Is a Transcriptional Corepressor , 1999, Molecular and Cellular Biology.

[62]  E. Olson,et al.  Association of Class II Histone Deacetylases with Heterochromatin Protein 1: Potential Role for Histone Methylation in Control of Muscle Differentiation , 2002, Molecular and Cellular Biology.