Although histone acetylation is one of the most widely studied epigenetic modifications, there is still a lack of information regarding how the acetylome is regulated during brain development and pathophysiological processes. We demonstrate that the embryonic brain (E15) is characterized by an increase in H3K9 acetylation as well as decreases in the levels of HDAC1 and HDAC3. Moreover, experimental induction of H3K9 hyperacetylation led to the overexpression of NCAM in the embryonic cortex and depletion of Sox2 in the subventricular ependyma, which mimicked the differentiation processes. Inducing differentiation in HDAC1-deficient mouse ESCs resulted in early H3K9 deacetylation, Sox2 downregulation, and enhanced astrogliogenesis, whereas neuro-differentiation was almost suppressed. Neuro-differentiation of (wt) ESCs was characterized by H3K9 hyperacetylation that was associated with HDAC1 and HDAC3 depletion. Conversely, the hippocampi of schizophrenia-like animals showed H3K9 deacetylation that was regulated by an increase in both HDAC1 and HDAC3. The hippocampi of schizophrenia-like brains that were treated with the cannabinoid receptor-1 inverse antagonist AM251 expressed H3K9ac at the level observed in normal brains. Together, the results indicate that co-regulation of H3K9ac by HDAC1 and HDAC3 is important to both embryonic brain development and neuro-differentiation as well as the pathophysiology of a schizophrenia-like phenotype.

• MeSH
• Acetylation MeSH
• Cannabinoid Receptor Antagonists pharmacology   MeSH
• Antipsychotic Agents pharmacology   MeSH
• Time Factors MeSH
• Epigenesis, Genetic MeSH
• Gestational Age MeSH
• Histone Deacetylase 1 antagonists & inhibitors   genetics   metabolism   MeSH
• Histone Deacetylases genetics   metabolism   MeSH
• Histones metabolism   MeSH
• Histone Deacetylase Inhibitors pharmacology   MeSH
• Methylazoxymethanol Acetate MeSH
• Disease Models, Animal MeSH
• Neural Cell Adhesion Molecules genetics   metabolism   MeSH
• Brain drug effects   embryology   enzymology   pathology   MeSH
• Mice, Inbred C57BL MeSH
• Neurogenesis * drug effects   MeSH
• Neurons drug effects   enzymology   pathology   MeSH
• Protein Processing, Post-Translational MeSH
• Rats, Sprague-Dawley MeSH
• Receptor, Cannabinoid, CB1 antagonists & inhibitors   metabolism   MeSH
• Schizophrenia chemically induced   drug therapy   enzymology   genetics   MeSH
• Signal Transduction MeSH
• SOXB1 Transcription Factors genetics   metabolism   MeSH
• Gene Expression Regulation, Developmental MeSH
• Animals MeSH
• Check Tag
• Animals MeSH
• Publication type
• Journal Article MeSH

• MeSH
• Astrocytoma enzymology   genetics   MeSH
• Cell Nucleus metabolism   MeSH
• Research Support as Topic MeSH
• Gliosis metabolism   MeSH
• Histone Deacetylases genetics   metabolism   MeSH
• Microscopy, Confocal MeSH
• Humans MeSH
• RNA, Messenger metabolism   MeSH
• Molecular Sequence Data MeSH
• Brain Neoplasms enzymology   genetics   MeSH
• Protein Isoforms genetics   metabolism   MeSH
• Gene Expression Regulation, Neoplastic MeSH
• Amino Acid Sequence MeSH
• Check Tag
• Humans MeSH

NKAP is a ubiquitously expressed nucleoplasmic protein that is currently known as a transcriptional regulatory molecule via its interaction with HDAC3 and spliceosomal proteins. Here, we report a disorder of transcriptional regulation due to missense mutations in the X chromosome gene, NKAP. These mutations are clustered in the C-terminal region of NKAP where NKAP interacts with HDAC3 and post-catalytic spliceosomal complex proteins. Consistent with a role for the C-terminal region of NKAP in embryogenesis, nkap mutant zebrafish with a C-terminally truncated NKAP demonstrate severe developmental defects. The clinical features of affected individuals are highly conserved and include developmental delay, hypotonia, joint contractures, behavioral abnormalities, Marfanoid habitus, and scoliosis. In affected cases, transcriptome analysis revealed the presence of a unique transcriptome signature, which is characterized by the downregulation of long genes with higher exon numbers. These observations indicate the critical role of NKAP in transcriptional regulation and demonstrate that perturbations of the C-terminal region lead to developmental defects in both humans and zebrafish.

• MeSH
• Zebrafish genetics   MeSH
• Down-Regulation genetics   MeSH
• Exons genetics   MeSH
• Transcription, Genetic genetics   MeSH
• Genes, X-Linked genetics   MeSH
• Histone Deacetylases genetics   MeSH
• Cognitive Dysfunction genetics   MeSH
• Humans MeSH
• Mutation, Missense genetics   MeSH
• Gene Expression Regulation genetics   MeSH
• Repressor Proteins genetics   MeSH
• Amino Acid Sequence MeSH
• Sequence Alignment MeSH
• Transcriptome genetics   MeSH
• Animals MeSH
• Check Tag
• Humans MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH

Signaling through the androgen receptor (AR) plays a critical role in prostate cancer progression. The AR is a classical nuclear receptor (NR) providing a link between signaling molecule and transcription response. Histone deacetylase inhibitors (HDACI) have antiproliferative and proapoptotic effects on prostate cancer cells and their implication in silence AR signaling may have potential therapeutic use. We aimed to study the inhibitory effects of the corepressor SMRT (Silencing Mediator for Retinoid and Thyroid hormone receptors) which forms a complex together with nuclear receptor corepressor (N-CoR) and with histone deacetylase 3 (HDAC3) on AR activity. The androgen-sensitive prostate cancer cell line LNCaP and androgen-insensitive prostate cancer cell line C4-2 both AR-positive, and androgen-insensitive DU145 and PC3 prostate cancer cell lines were treated with two HDACIs, sodium butyrate (NaB) and/or trichostatin A (TSA). We amplified immunoprecipitated DNA by conventional PCR and in the following step we used the chromatin immunoprecipitation (ChIP) analysis coupled with quantitative PCR for monitoring NaB induced formation of AR-SMRT/N-CoR complex binding on the PSA promoter. The co-immunoprecipitation assay revealed increase in AR-SMRT formation in NaB treated cells. Simultaneously, the Western blot analysis showed a significant decrease in AR protein expression. Furthermore, we estimated the reduced presence of HDAC2 and HDAC3 proteins by NaB and TSA treatment in AR-negative DU145 cell line. In conclusion, the inhibitory effect of NaB on AR gene expression seems to be specific and unique for prostate cancer AR-positive cell lines and corresponds with its ability to stimulate AR-SMRT complex formation. We suggest that AR and SMRT/N-CoR corepressors may form a stable complex in vitro and NaB may facilitate the interaction between AR nuclear steroid receptor and SMRT corepressor protein.

• MeSH
• Receptors, Androgen genetics   metabolism   MeSH
• Butyrates metabolism   pharmacology   therapeutic use   MeSH
• Time Factors MeSH
• Histone Deacetylase 2 metabolism   MeSH
• Histone Deacetylases metabolism   MeSH
• Immunoprecipitation MeSH
• Histone Deacetylase Inhibitors therapeutic use   MeSH
• Nuclear Receptor Co-Repressor 2 genetics   metabolism   MeSH
• Hydroxamic Acids metabolism   pharmacology   therapeutic use   MeSH
• Humans MeSH
• Cell Line, Tumor MeSH
• Prostatic Neoplasms drug therapy   genetics   metabolism   MeSH
• Gene Expression Regulation, Neoplastic MeSH
• Protein Binding genetics   MeSH
• Dose-Response Relationship, Drug MeSH
• Check Tag
• Humans MeSH
• Male MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH
• Comparative Study MeSH

Zinc-dependent histone deacetylases (HDACs) and sirtuins (SIRT) represent two different classes of enzymes which are responsible for deacylation of modified lysine side chains. The repertoire of acyl residues on lysine side chains identified in vivo is rapidly growing, and very recently lysine lactoylation was described to be involved in metabolic reprogramming. Additionally, lysine pyruvoylation represents a marker for aging and liver cirrhosis. Here, we report a systematic analysis of acyl-specificity of human zinc-dependent HDAC and sirtuin isoforms. We identified HDAC3 as a robust delactoylase with several-thousand-fold higher activity as compared to SIRT2, which was claimed to be the major in vivo delactoylase. Additionally, we systematically searched for enzymes, capable of removing pyruvoyl residues from lysine side chains. Using model peptides, we uncovered high depyruvoylase activity for HDAC6 and HDAC8. Interestingly, such substrates have extremely low KM values for both HDAC isoforms, pointing to possible in vivo functions.

• MeSH
• Histone Deacetylases MeSH
• Histone Deacetylase Inhibitors MeSH
• Humans MeSH
• Lysine * chemistry   MeSH
• Protein Isoforms MeSH
• Repressor Proteins metabolism   MeSH
• Sirtuin 2 * MeSH
• Aging MeSH
• Zinc MeSH
• Check Tag
• Humans MeSH
• Publication type
• Journal Article MeSH
• Systematic Review MeSH

Class I histone deacetylases (HDACs) are key regulators of cell proliferation and they are frequently dysregulated in cancer cells. We report here the synthesis of a novel series of class-I selective HDAC inhibitors (HDACi) containing a 2-aminobenzamide moiety as a zinc-binding group connected with a central (piperazin-1-yl)pyrazine or (piperazin-1-yl)pyrimidine moiety. Some of the compounds were additionally substituted with an aromatic capping group. Compounds were tested in vitro against human HDAC1, 2, 3, and 8 enzymes and compared to reference class I HDACi (Entinostat (MS-275), Mocetinostat, CI994 and RGFP-966). The most promising compounds were found to be highly selective against HDAC1, 2 and 3 over the remaining HDAC subtypes from other classes. Molecular docking studies and MD simulations were performed to rationalize the in vitro data and to deduce a complete structure activity relationship (SAR) analysis of this novel series of class-I HDACi. The most potent compounds, including 19f, which blocks HDAC1, HDAC2, and HDAC3, as well as the selective HDAC1/HDAC2 inhibitors 21a and 29b, were selected for further cellular testing against human acute myeloid leukemia (AML) and erythroleukemic cancer (HEL) cells, taking into consideration their low toxicity against human embryonic HEK293 cells. We found that 19f is superior to the clinically tested class-I HDACi Entinostat (MS-275). Thus, 19f is a new and specific HDACi with the potential to eliminate blood cancer cells of various origins.

• MeSH
• Benzamides chemical synthesis   chemistry   pharmacology   MeSH
• HEK293 Cells MeSH
• Histone Deacetylase Inhibitors chemical synthesis   chemistry   pharmacokinetics   pharmacology   MeSH
• Humans MeSH
• Cell Line, Tumor MeSH
• ortho-Aminobenzoates chemical synthesis   chemistry   MeSH
• Antineoplastic Agents chemical synthesis   chemistry   pharmacokinetics   pharmacology   MeSH
• Proton Magnetic Resonance Spectroscopy MeSH
• Pyrazines chemistry   MeSH
• Pyridines chemical synthesis   chemistry   pharmacology   MeSH
• Molecular Docking Simulation * MeSH
• Check Tag
• Humans MeSH
• Publication type
• Journal Article MeSH

We studied the histone signature of embryonic and adult brains to strengthen existing evidence of the importance of the histone code in mouse brain development. We analyzed the levels and distribution patterns of H3K9me1, H3K9me2, H3K9me3, and HP1β in both embryonic and adult brains. Western blotting showed that during mouse brain development, the levels of H3K9me1, H3K9me2, and HP1β exhibited almost identical trends, with the highest protein levels occurring at E15 stage. These trends differed from the relatively stable level of H3K9me3 at developmental stages E8, E13, E15, and E18. Compared with embryonic brains, adult brains were characterized by very low levels of H3K9me1/me2/me3 and HP1β. Manipulation of the embryonic epigenome through histone deacetylase inhibitor treatment did not affect the distribution patterns of the studied histone markers in embryonic ventricular ependyma. Similarly, Hdac3 depletion in adult animals had no effect on histone methylation in the adult hippocampus. Our results indicate that the distribution of HP1β in the embryonic mouse brain is related to that of H3K9me1/me2 but not to that of H3K9me3. The unique status of H3K9me3 in the brain was confirmed by its pronounced accumulation in the granular layer of the adult olfactory bulb. Moreover, among the studied proteins, H3K9me3 was the only posttranslational histone modification that was highly abundant at clusters of centromeric heterochromatin, called chromocenters. When we focused on the hippocampus, we found this region to be rich in H3K9me1 and H3K9me3, whereas H3K9me2 and HP1β were present at a very low level or even absent in the hippocampal blade. Taken together, these results revealed differences in the epigenome of the embryonic and adult mouse brain and showed that the adult hippocampus, the granular layer of the adult olfactory bulb, and the ventricular ependyma of the embryonic brain are colonized by specific epigenetic marks.

• MeSH
• Chromosomal Proteins, Non-Histone analysis   metabolism   MeSH
• Microscopy, Fluorescence MeSH
• Histone-Lysine N-Methyltransferase metabolism   MeSH
• Immunohistochemistry MeSH
• Brain embryology   metabolism   MeSH
• Mice, Inbred C57BL MeSH
• Mice, Knockout MeSH
• Mice MeSH
• Animals MeSH
• Check Tag
• Mice MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH

Signaling through the androgen receptor (AR) plays a critical role in prostate cancer progression. The AR is a classical nuclear receptor (NR) providing a link between signaling molecule and transcription response. Histone deacetylase inhibitors- (HDACI) have antiproliferative and proapoptotic effects on prostate cancer cells and their implication in silence AR signaling may have potential therapeutic use. We aimed to study the inhibitory effects of the corepressor SMRT (Silencing Mediator for Retinoid and Thyroid -hormone receptors) which forms a complex together with nuclear receptor corepressor (N-CoR) and with histone deacetylase 3 (HDAC3) on AR activity.The androgen-sensitive prostate cancer cell line LNCaP and androgen-insensitive prostate cancer cell line C4-2 both AR-positive, and androgen-insensitive DU145 and PC3 prostate cancer cell lines were treated with two HDACIs, sodium butyrate (NaB) and/or trichostatin A (TSA). We amplified immunoprecipitated DNA by conventional PCR and in the -following step we used the chromatin immunoprecipitation (ChIP) analysis coupled with quantitative PCR for monitoring NaB induced formation of AR-SMRT/N-CoR complex binding on the PSA promoter. The co-immunoprecipitation assay revealed increase in AR-SMRT formation in NaB treated cells. Simultaneously, the Western blot analysis showed a significant decrease in AR protein expression. In conclusion, the inhibitory effect of NaB on AR gene expression seems to be specific and unique for prostate cancer AR-positive cell lines and corresponds with its ability to stimulate AR-SMRT complex formation. We suggest that AR and SMRT/N-CoR corepressors may form a stable complex in vitro and NaB may facilitate the interaction between AR nuclear steroid receptor and SMRT corepressor prote.

• MeSH
• Receptors, Androgen metabolism   MeSH
• Butyrates pharmacology   MeSH
• Histone Deacetylase 2 analysis   MeSH
• Histone Deacetylases analysis   metabolism   MeSH
• Histone Deacetylase Inhibitors therapeutic use   MeSH
• Nuclear Receptor Co-Repressor 1 metabolism   MeSH
• Hydroxamic Acids pharmacology   MeSH
• Humans MeSH
• Cell Line, Tumor MeSH
• Prostatic Neoplasms drug therapy   metabolism   MeSH
• Promoter Regions, Genetic MeSH
• Prostate-Specific Antigen genetics   MeSH
• Multidrug Resistance-Associated Proteins metabolism   MeSH
• Check Tag
• Humans MeSH
• Male MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH