Maternal histone deacetylase is accumulated in the nuclei of Xenopus oocytes as protein complexes with potential enzyme activity

1999 ◽  
Vol 112 (14) ◽  
pp. 2441-2452 ◽  
Author(s):  
J. Ryan ◽  
A.J. Llinas ◽  
D.A. White ◽  
B.M. Turner ◽  
J. Sommerville
Keyword(s):  
Histone Deacetylases ◽  
Protein Complexes ◽  
Trichostatin A ◽  
Hdac Activity ◽  
Nuclear Complex ◽  
Phosphatase Treatment ◽  
Core Histones ◽  
Histone Deposition ◽  
Potential Enzyme Activity ◽  
Transcription And Replication

Reversible acetylation of core histones plays an important regulatory role in transcription and replication of chromatin. The acetylation status of chromatin is determined by the equilibrium between activities of histone acetyltransferases (HATs) and histone deacetylases (HDACs). The Xenopus protein HDACm shows sequence homology to other putative histone deacetylases, but its mRNA is expressed only during early development. Both HDACm protein and acetylated non-chromosomal histones are accumulated in developing oocytes, indicating that the key components for histone deposition into new chromatin during blastula formation are in place by the end of oogenesis. Here we show that the 57 kDa HDACm protein undergoes steady accumulation in the nucleus, where it is organized in a multiprotein complex of approx. 300 kDa. A second, major component of the nuclear complex is the retinoblastoma-associated protein p48 (RbAp48/46), which may be used as an adaptor to contact acetylated histones in newly assembled chromatin. The nuclear complex has HDAC activity that is sensitive to trichostatin A, zinc ions and phosphatase treatment. The 57 kDa protein serves as a marker for total HDAC activity throughout oogenesis and early embryogenesis. The active HDACm complex and its acetylated histone substrates appear to be kept apart until after chromatin assembly has taken place. However, recombinant HDACm, injected into the cytoplasm of oocytes, not only is translocated to the nucleus, but also is free to interact with the endogenous chromatin.

scholarly journals Nkx3.1 binds and negatively regulates the transcriptional activity of Sp-family members in prostate-derived cells

2005 ◽  
Vol 393 (1) ◽  
pp. 397-409 ◽  
Author(s):  
Steven O. Simmons ◽  
Jonathan M. Horowitz
Keyword(s):  
Dna Binding ◽  
Histone Deacetylases ◽  
Target Genes ◽  
Protein Complexes ◽  
Trichostatin A ◽  
Prostate Gland ◽  
Specific Antigen ◽  
Family Members ◽  
Early Event ◽  
Prostate Tumorigenesis

Nkx3.1 is a homeodomain-containing transcription factor that is expressed early in the development of the prostate gland and is believed to play an important role in the differentiation of prostatic epithelia. Loss of Nkx3.1 protein expression is often an early event in prostate tumorigenesis, and the abundance of Nkx3.1-negative epithelial cells increases with disease progression. In a number of systems, homeodomain proteins collaborate with zinc-finger-containing transcription factors to bind and regulate target genes. In the present paper, we report that Nkx3.1 collaborates with Sp-family members in the regulation of PSA (prostate-specific antigen) in prostate-derived cells. Nkx3.1 forms protein complexes with Sp proteins that are dependent on their respective DNA-binding domains and an N-terminal segment of Nkx3.1, and Nkx3.1 negatively regulates Sp-mediated transcription via Trichostatin A-sensitive and -insensitive mechanisms. A distal 1000 bp portion of the PSA promoter is required for transrepression by Nkx3.1, although Nkx3.1 DNA-binding activity is itself not required. We conclude that Nkx3.1 negatively regulates Sp-mediated transcription via the tethering of histone deacetylases and/or by inhibiting the association of Sp proteins with co-activators.

NF-κB inhibits transcription of the H+-K+-ATPase α2-subunit gene: role of histone deacetylases

2002 ◽  
Vol 283 (5) ◽  
pp. F904-F911 ◽  
Author(s):  
Wenzheng Zhang ◽  
Bruce C. Kone
Keyword(s):  
Histone Deacetylases ◽  
Transcriptional Control ◽  
Protein Complexes ◽  
Trichostatin A ◽  
Collecting Duct ◽  
Proximal Promoter ◽  
Inner Medullary Collecting Duct ◽  
Nuclear Extracts ◽  
Medullary Collecting Duct

The H+-K+-ATPase α2 (HKα2) gene plays a central role in potassium homeostasis, yet little is known about its transcriptional control. We recently demonstrated that the proximal promoter confers basal transcriptional activity in mouse inner medullary collecting duct 3 cells. We sought to determine whether the κB DNA binding element at −104 to −94 influences basal HKα2 gene transcription in these cells. Recombinant NF-κB p50 footprinted the region −116/−94 in vitro. Gel shift and supershift analysis revealed NF-κB p50- and p65-containing DNA-protein complexes in nuclear extracts of mouse inner medullary collecting duct 3 cells. A promoter-luciferase construct with a mutated −104/−94 NF-κB element exhibited higher activity than the wild-type promoter in transfection assays. Overexpression of NF-κB p50, p65, or their combination trans-repressed the HKα2 promoter. The histone deacetylase (HDAC) inhibitor trichostatin A partially reversed NF-κB-mediated trans-repression of the HKα2 promoter. HDAC6 overexpression inhibited HKα2 promoter activity, and HDAC6 coimmunoprecipitated with NF-κB p50 and p65. These results suggest that HDAC6, recruited to the DNA protein complex, acts with NF-κB to suppress HKα2 transcription and identify NF-κB p50 and p65 as novel binding partners for HDAC6.

Competitive inhibition of histone deacetylase activity by trichostatin A and butyrate

2007 ◽  
Vol 85 (6) ◽  
pp. 751-758 ◽  
Author(s):  
Anoushe Sekhavat ◽  
Jian-Min Sun ◽  
James R. Davie
Keyword(s):  
Histone Deacetylases ◽  
Competitive Inhibition ◽  
Trichostatin A ◽  
Hdac Inhibitors ◽  
Cancer Cell Line ◽  
Kinetic Studies ◽  
Competitive Inhibitor ◽  
Hdac Activity ◽  
Modeling Data ◽  
Sin3 Complex

Histone deacetylases (HDACs) play a pivotal role in gene expression through their involvement in chromatin remodeling. The abnormal targeting or retention of HDACs to DNA regulatory regions is observed in many cancers, and hence HDAC inhibitors are being tested as promising anti-tumor agents. The results of previous kinetic studies, characterizing trichostatin A (TSA), as well as butyrate, as HDAC noncompetitive inhibitors, conflict with crystallographic and homology modeling data suggesting that TSA should act as a competitive inhibitor. Our results demonstrate that each of the HDAC inhibitors TSA and butyrate inhibits HDAC activity in a competitive fashion. Co-immunoprecipitation studies show that the inhibition of HDAC1 and HDAC2 activity by TSA does not disturb the extensive level of their association in the human breast cancer cell line MCF-7. Moreover, the inhibition of HDAC activity by TSA does not interfere with the interaction of HDAC1 and HDAC2 with Sin3A, a core component of the Sin3 complex. Thus, repressor complexes such as Sin3, appear to be stable in the presence of TSA. The association of HDAC2 with transcription factor Sp1 is also not affected by TSA.

scholarly journals Sepsis and glucocorticoids upregulate p300 and downregulate HDAC6 expression and activity in skeletal muscle

2010 ◽  
Vol 299 (2) ◽  
pp. R509-R520 ◽  
Author(s):  
Nima Alamdari ◽  
Ira J. Smith ◽  
Zaira Aversa ◽  
Per-Olof Hasselgren
Keyword(s):  
Skeletal Muscle ◽  
Histone Deacetylases ◽  
Muscle Wasting ◽  
Trichostatin A ◽  
Histone Acetyl Transferase ◽  
Hdac Activity ◽  
Protein Levels ◽  
Expression And Activity

Muscle wasting during sepsis is in part regulated by glucocorticoids. In recent studies, treatment of cultured muscle cells in vitro with dexamethasone upregulated expression and activity of p300, a histone acetyl transferase (HAT), and reduced expression and activity of the histone deacetylases-3 (HDAC3) and -6, changes that favor hyperacetylation. Here, we tested the hypothesis that sepsis and glucocorticoids regulate p300 and HDAC3 and -6 in skeletal muscle in vivo. Because sepsis-induced metabolic changes are particularly pronounced in white, fast-twitch skeletal muscle, most experiments were performed in extensor digitorum longus muscles. Sepsis in rats upregulated p300 mRNA and protein levels, stimulated HAT activity, and reduced HDAC6 expression and HDAC activity. The sepsis-induced changes in p300 and HDAC expression were prevented by the glucocorticoid receptor antagonist RU38486. Treatment of rats with dexamethasone increased expression of p300 and HAT activity, reduced expression of HDAC3 and -6, and inhibited HDAC activity. Finally, treatment with the HDAC inhibitor trichostatin A resulted in increased muscle proteolysis and expression of the ubiquitin ligase atrogin-1. Taken together, our results suggest for the first time that sepsis-induced muscle wasting may be regulated by glucocorticoid-dependent hyperacetylation caused by increased p300 and reduced HDAC expression and activity. The recent development of pharmacological HDAC activators may provide a novel avenue to prevent and treat muscle wasting in sepsis and other catabolic conditions.

scholarly journals HdaA, a Major Class 2 Histone Deacetylase of Aspergillus nidulans, Affects Growth under Conditions of Oxidative Stress

2005 ◽  
Vol 4 (10) ◽  
pp. 1736-1745 ◽  
Author(s):  
Martin Tribus ◽  
Johannes Galehr ◽  
Patrick Trojer ◽  
Gerald Brosch ◽  
Peter Loidl ◽  
...  
Keyword(s):  
Oxidative Stress ◽  
Aspergillus Nidulans ◽  
Histone Deacetylases ◽  
Expression System ◽  
Intrinsic Activity ◽  
Regulation Of Transcription ◽  
Hdac Activity ◽  
Amino Terminal ◽  
Prokaryotic Expression System ◽  
Core Histones

ABSTRACT Histone deacetylases (HDACs) catalyze the removal of acetyl groups from the ε-amino group of distinct lysine residues in the amino-terminal tail of core histones. Since the acetylation status of core histones plays a crucial role in fundamental processes in eukaryotic organisms, such as replication and regulation of transcription, recent research has focused on the enzymes responsible for the acetylation/deacetylation of core histones. Very recently, we showed that HdaA, a member of the Saccharomyces cerevisiae HDA1-type histone deacetylases, is a substantial contributor to total HDAC activity in the filamentous fungus Aspergillus nidulans. Now we demonstrate that deletion of the hdaA gene indeed results in the loss of the main activity peak and in a dramatic reduction of total HDAC activity. In contrast to its orthologs in yeast and higher eukaryotes, HdaA has strong intrinsic activity as a protein monomer when expressed as a recombinant protein in a prokaryotic expression system. In vivo, HdaA is involved in the regulation of enzymes which are of vital importance for the cellular antioxidant response in A. nidulans. Consequently, ΔhdaA strains exhibit significantly reduced growth on substrates whose catabolism generates molecules responsible for oxidative stress conditions in the fungus. Our analysis revealed that reduced expression of the fungal catalase CatB is jointly responsible for the significant growth reduction of the hdaA mutant strains.

scholarly journals Cloning and characterization of a novel human histone deacetylase, HDAC8

2000 ◽  
Vol 350 (1) ◽  
pp. 199-205 ◽  
Author(s):  
Joseph J. BUGGY ◽  
Mindy L. SIDERIS ◽  
Polly MAK ◽  
Donald D. LORIMER ◽  
Brian MCINTOSH ◽  
...  
Keyword(s):  
Histone Deacetylases ◽  
Cell Transformation ◽  
Trichostatin A ◽  
Protein Product ◽  
Histone H4 ◽  
Hybrid Mapping ◽  
Peptide Substrates ◽  
Acetylation State ◽  
Core Histones

Histone deacetylases (HDACs) are a growing family of enzymes implicated in transcriptional regulation by affecting the acetylation state of core histones in the nucleus of cells. HDACs are known to have key roles in the regulation of cell proliferation [Brehm, Miska, McCance, Reid, Bannister and Kouzarides (1998) Nature (London) 391, 597–600], and aberrant recruitment of an HDAC complex has been shown to be a key step in the mechanism of cell transformation in acute promyelocytic leukaemia [Grignani, De Matteis, Nervi, Tomassoni, Gelmetti, Cioce, Fanelli, Ruthardt, Ferrara, Zamir et al. (1998) Nature (London) 391, 815–818; Lin, Nagy, Inoue, Shao, Miller and Evans (1998), Nature (London) 391, 811–814]. Here we present the complete nucleotide sequence of a cDNA clone, termed HDAC8, that encodes a protein product with similarity to the RPD3 class (I) of HDACs. The predicted 377-residue HDAC8 product contains a shorter C-terminal extension relative to other members of its class. After expression in two cell systems, immunopurified HDAC8 is shown to possess trichostatin A- and sodium butyrate-inhibitable HDAC activity on histone H4 peptide substrates as well as on core histones. Expression profiling reveals the expression of HDAC8 to various degrees in every tissue tested and also in several tumour lines. Mutation of two adjacent histidine residues within the predicted active site severely decreases activity, confirming these residues as important for HDAC8 enzyme activity. Finally, linkage analysis after radiation hybrid mapping has localized HDAC8 to chromosomal position Xq21.2–Xq21.3. These results confirm HDAC8 as a new member of the HDAC family.

Abstract 654: Overexpression Of Class I Histone Deacetylases Repress Npr1 Gene Transcription In Mouse Mesangial Cells

Hypertension ◽  
2012 ◽  
Vol 60 (suppl_1) ◽  
Author(s):  
Prerna Kumar ◽  
Kailash N Pandey
Keyword(s):  
Gene Transcription ◽  
Hdac Inhibitor ◽  
Histone Deacetylases ◽  
Promoter Activity ◽  
Mesangial Cells ◽  
Trichostatin A ◽  
Class I ◽  
Mrna Levels ◽  
Signaling Mechanism ◽  
Hdac Activity

Activation of guanylyl cyclase/natriuretic peptide receptor-A (GC-A/NPRA) by cardiac hormones atrial and brain natriuretic peptides (ANP and BNP) produces the second messenger cGMP, which plays a pivotal role in maintaining blood pressure and cardiovascular homeostasis. The objective of the present study was to gain insight into the epigenetic signaling mechanism in the regulation of Npr1 (coding for GC-A/NPRA) gene transcription and expression. The mouse mesangial cells were cultured in Dulbecco’s modified Eagles medium containing 10% fetal bovine serum and ITS (insulin, transferrin, and sodium selenite). The results showed that class I and II histone deacetylase (HDAC) inhibitors trichostatin A (TSA) and sodium butyrate (NaBu) significantly induced Npr1 promoter activity by 5- and 7-fold (TSA-treated, 2160.3 ± 32.4 and NaBu-treated, 3024.89 ± 25.03 vs. untreated control 432.2 ± 23.2, p < 0.001). TSA induced Npr1 mRNA levels by 3.5-fold and NaBu by 5-fold (TSA-treated, 9.7 ± 0.89 and NaBu-treated, 13.8 ± 1.9 vs. control 2.68 ± 0.5; p < 0.001). NaBu significantly reduced HDAC activity by 50% (5.15 ± 0.11 vs. control 1.94 ± 0.26) and stimulated histone acetyltransferase activity by 3-fold (48.93 ± 7.32 vs. control 15.98 ± 2.1). Furthermore, class I specific HDAC inhibitor mocetinostat (MGCD0103) significantly enhanced Npr1 promoter activity by 6-fold (1955.4 ± 47.32 vs. control 325.9 ± 23.4, p < 0.001) whereas class II HDAC inhibitor MC 1568 showed no significant effect on the promoter activity. MGCD0103 increased the acetylation of histones H3 by 2.5-fold (8850 ± 90.4 vs. control 3421 ± 68.5) and H4 by 4-fold (27320 ± 213 vs. control 6919 ± 78.5), respectively. Overexpression of class I HDACs (HDAC1 and HDAC2) repressed Npr1 promoter activity by 50% (234.8 ± 9.5 and 265.4 ± 10.32, respectively vs. control 543.23 ± 7.8, p < 0.001) and knockdown of HDAC1 and HDAC2 expression by siRNA abolished this repression. Collectively, the present results demonstrate that class I HDAC inhibitor upregulates Npr1 gene transcription via inhibition of HDAC activity and histone acetylation. The identification of epigenetic signaling as a regulator of Npr1 gene expression will have important implications in prevention of hypertension and cardiac remodeling.

A Homogeneous Nonisotopic Histone Deacetylase Activity Assay

2003 ◽  
Vol 8 (1) ◽  
pp. 89-95 ◽  
Author(s):  
Birgit Heltweg ◽  
Manfred Jung
Keyword(s):  
Drug Discovery ◽  
Fluorescence Quenching ◽  
Histone Deacetylase ◽  
Anticancer Agents ◽  
Histone Deacetylases ◽  
Activity Assay ◽  
Animal Testing ◽  
Hdac Activity

Histone deacetylases (HDACs) are important regulators of transcription, and their inhibitors are a promising class of anticancer agents. The methods for the determination of HDAC activity and its inhibition that are currently available suffer from various drawbacks, such as animal testing, radioactive substrates, or limited throughput. Therefore, a fast nonisotopic method for the measurement of HDAC activity is highly desirable. The authors present such an assay that relies on the fluorescent HDAC substrate developed previously in their group. After incubation of the substrate with the enzyme, a derivatization leads to efficient fluorescence quenching in the deacetylated metabolite. Thus, only the fluorescence emitted by the remaining substrate is detected, which allows for a convenient detection of HDAC activity in a homogeneous format that can be performed on multiwell plate readers. This procedure, called HDASH (histone deacetylase assay—homogeneous), should be a valuable tool in transcriptional research and especially drug discovery. ( Journal of Biomolecular Screening 2003:89-95)

α2-Adrenoceptor agonist dexmedetomidine protects septic acute kidney injury through increasing BMP-7 and inhibiting HDAC2 and HDAC5

2012 ◽  
Vol 303 (10) ◽  
pp. F1443-F1453 ◽  
Author(s):  
Chung-Hsi Hsing ◽  
Chiou-Feng Lin ◽  
Edmund So ◽  
Ding-Ping Sun ◽  
Tai-Chi Chen ◽  
...  
Keyword(s):  
Acute Kidney Injury ◽  
Histone Deacetylases ◽  
Trichostatin A ◽  
Histone H3 ◽  
Cecal Ligation ◽  
Kidney Injury ◽  
Protective Effects ◽  
Tnf Α

Bone morphogenetic protein (BMP)-7 protects sepsis-induced acute kidney injury (AKI). Dexmedetomidine (DEX), an α2-adrenoceptor (α2-AR) agonist, has anti-inflammatory effects. We investigated the protective effects of DEX on sepsis-induced AKI and the expression of BMP-7 and histone deacetylases (HDACs). In vitro , the effects of DEX or trichostatin A (TSA, an HDAC inhibitor) on TNF-α, monocyte chemotactic protein (MCP-1), BMP-7, and HDAC mRNA expression in LPS-stimulated rat renal tubular epithelial NRK52E cells, was determined using real-time PCR. In vivo, mice were intraperitoneally injected with DEX (25 μg/kg) or saline immediately and 12 h after cecal ligation and puncture (CLP) surgery. Twenty-four hours after CLP, we examined kidney injury and renal TNF-α, MCP-1, BMP-7, and HDAC expression. Survival was monitored for 120 h. LPS increased HDAC2, HDAC5, TNF-α, and MCP-1 expression, but decreased BMP-7 expression in NRK52E cells. DEX treatment decreased the HDAC2, HDAC5, TNF-α, and MCP-1 expression, but increased BMP-7 and acetyl histone H3 expression, whose effects were blocked by yohimbine, an α2-AR antagonist. With DEX treatment, the LPS-induced TNF-α expression and cell death were attenuated in scRNAi-NRK52E but not BMP-7 RNAi-NRK52E cells. In CLP mice, DEX treatment increased survival and attenuated AKI. The expression of HDAC2, HDAC5, TNF-α, and MCP-1 mRNA in the kidneys of CLP mice was increased, but BMP-7 was decreased. However, DEX treatment reduced those changes. DEX reduces sepsis-induced AKI by decreasing TNF-α and MCP-1 and increasing BMP-7, which is associated with decreasing HDAC2 and HDAC5, as well as increasing acetyl histone H3.

scholarly journals Deacetylation of Histone H4 Accompanying Cardiomyogenesis is Weakened in HDAC1-Depleted ES Cells

2018 ◽  
Vol 19 (8) ◽  
pp. 2425 ◽  
Author(s):  
Orazio Angelo Arcidiacono ◽  
Jana Krejčí ◽  
Jana Suchánková ◽  
Eva Bártová
Keyword(s):  
Histone Deacetylases ◽  
Trichostatin A ◽  
Es Cells ◽  
Embryonic Stem ◽  
Cardiomyocyte Differentiation ◽  
Histone H4 ◽  
Epigenetic Factors ◽  
Double Knockout ◽  
Physiological Processes

Cell differentiation into cardiomyocytes requires activation of differentiation-specific genes and epigenetic factors that contribute to these physiological processes. This study is focused on the in vitro differentiation of mouse embryonic stem cells (mESCs) induced into cardiomyocytes. The effects of clinically promising inhibitors of histone deacetylases (HDACi) on mESC cardiomyogenesis and on explanted embryonic hearts were also analyzed. HDAC1 depletion caused early beating of cardiomyocytes compared with those of the wild-type (wt) counterpart. Moreover, the adherence of embryonic bodies (EBs) was reduced in HDAC1 double knockout (dn) mESCs. The most important finding was differentiation-specific H4 deacetylation observed during cardiomyocyte differentiation of wt mESCs, while H4 deacetylation was weakened in HDAC1-depleted cells induced to the cardiac pathway. Analysis of the effect of HDACi showed that Trichostatin A (TSA) is a strong hyperacetylating agent, especially in wt mESCs, but only SAHA reduced the size of the beating areas in EBs that originated from HDAC1 dn mESCs. Additionally, explanted embryonic hearts (e15) responded to treatment with HDACi: all of the tested HDACi (TSA, SAHA, VPA) increased the levels of H3K9ac, H4ac, H4K20ac, and pan-acetylated lysines in embryonic hearts. This observation shows that explanted tissue can be maintained in a hyperacetylation state several hours after excision, which appears to be useful information from the view of transplantation strategy and the maintenance of gene upregulation via acetylation in tissue intended for transplantation.

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