Histone acetylation modulates alternative splicing of several hundred genes. Here, we tested the role of the histone acetyltransferase p300 in alternative splicing and showed that knockdown of p300 promotes inclusion of the fibronectin (FN1) alternative EDB exon. p300 associates with CRE sites in the promoter via the CREB transcription factor. We created mini-gene reporters driven by an artificial promoter containing CRE sites. Both deletion and mutation of the CRE site affected EDB alternative splicing in the same manner as p300 knockdown. Next we showed that p300 controls histone H4 acetylation along the FN1 gene. Consistently, p300 depletion and CRE deletion/mutation both reduced histone H4 acetylation on mini-gene reporters. Finally, we provide evidence that the effect of CRE inactivation on H4 acetylation and alternative splicing is counteracted by the inhibition of histone deacetylases. Together, these data suggest that histone acetylation could be one of the mechanisms how promoter and promoter binding proteins influence alternative splicing.

• Klíčová slova
• alternative splicing, fibronectin, histone acetylation, p300, promoter,
• MeSH
• acetylace MeSH
• alternativní sestřih * MeSH
• fibronektiny genetika   metabolismus   MeSH
• genový knockdown MeSH
• HeLa buňky MeSH
• histony metabolismus   MeSH
• integrasy genetika   MeSH
• lidé MeSH
• messenger RNA metabolismus   MeSH
• promotorové oblasti (genetika) MeSH
• protein p300 asociovaný s E1A genetika   metabolismus   MeSH
• reportérové geny MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• Cre recombinase MeSH Prohlížeč
• EP300 protein, human MeSH Prohlížeč
• fibronektiny MeSH
• FN1 protein, human MeSH Prohlížeč
• histony MeSH
• integrasy MeSH
• messenger RNA MeSH
• protein p300 asociovaný s E1A MeSH

Delayed fertilization leads to the ageing of post-ovulatory oocytes and reduces the developmental competence of arising embryos. Little information is available about the molecular processes during fish oocyte ageing. The current study investigated the functional consequences of oocyte ageing in grass carp Ctenopharyngodon idella embryos. In addition, the dynamics of selected post-transcriptionally modified histones (acetylation of H3K9, H3K14, H4K5, H4K8, H4K12, and H4K16) were analyzed during oocyte ageing. Ovulated oocytes were aged in vitro for 4 h in the laboratory incubator at 20 °C and studied for selected post-translational modification of histones. In addition, histone acetyltransferase activity was investigated as an important regulator of histone acetylation modification. The results indicated a significant decrease in oocyte fertilizing ability through 1 h of post-ovulatory ageing, and a complete loss of egg fertilizing abilities was detected at 4-h aged oocytes. Furthermore, post-ovulatory oocyte ageing for 1 and 4 h led to decreased levels of H4K12 acetylation. The activity of histone acetyltransferases increased significantly after ageing of the oocytes for 30 h in vitro. This modification may partly contribute to explaining the failures of egg viability and embryo development in the offspring from the aged oocytes. The results are the first to report histone modifications as a crucial epigenetic regulator during oocyte ageing in fish and might also benefit other vertebrates.

• Klíčová slova
• Ctenopharyngodon Idella, Epigenetics, Fertilization, HAT activity, Histone acetylation, Ovulation,
• MeSH
• acetylace MeSH
• histonacetyltransferasy * metabolismus   genetika   MeSH
• histony * metabolismus   MeSH
• kapři * metabolismus   MeSH
• oocyty * MeSH
• posttranslační úpravy proteinů MeSH
• stárnutí buněk MeSH
• zvířata MeSH
• Check Tag
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• histonacetyltransferasy * MeSH
• histony * MeSH

Aging is the most critical factor that influences the quality of post-ovulatory oocytes. Age-related molecular pathways remain poorly understood in fish oocytes. In this study, we examined the effect of oocyte aging on specific histone acetylation in common carp Cyprinus carpio. The capacity to progress to the larval stage in oocytes that were aged for 28 h in vivo and in vitro was evaluated. Global histone modifications and specific histone acetylation (H3K9ac, H3K14ac, H4K5ac, H4K8ac, H4K12ac, and H4K16ac) were investigated during oocyte aging. Furthermore, the activity of histone acetyltransferase (HAT) was assessed in fresh and aged oocytes. Global histone modifications did not exhibit significant alterations during 8 h of oocyte aging. Among the selected modifications, H4K12ac increased significantly at 28 h post-stripping (HPS). Although not significantly different, HAT activity exhibited an upward trend during oocyte aging. Results of our current study indicate that aging of common carp oocytes for 12 h results in complete loss of egg viability rates without any consequence in global and specific histone modifications. However, aging oocytes for 28 h led to increased H4K12ac. Thus, histone acetylation modification as a crucial epigenetic mediator may be associated with age-related defects, particularly in oocytes of a more advanced age.

• Klíčová slova
• Cyprinus carpio, egg quality, epigenetics, histone acetyltransferase, histone modifications, post-ovulatory aging,
• MeSH
• acetylace MeSH
• histonacetyltransferasy genetika   MeSH
• histony genetika   MeSH
• kapři genetika   růst a vývoj   MeSH
• oocyty růst a vývoj   metabolismus   MeSH
• posttranslační úpravy proteinů genetika   MeSH
• stárnutí genetika   MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• histonacetyltransferasy MeSH
• histony MeSH

Histone deacetylases (HDACs) target acetylated lysine residues in histone and non-histone proteins. HDACs are implicated in the regulation of genomic stability, cell cycle, cell death and differentiation and thus critically involved in tumorigenesis. Further, HDACs regulate T-cell development and HDAC inhibitors (HDACis) have been approved for clinical use in some T-cell malignancies. Still, the exact targets and mechanisms of HDAC inhibition in cancer are understudied. We isolated tumor cell lines from a transgenic mouse model of anaplastic large cell lymphoma (ALCL), a rare T-cell lymphoma, and abrogated HDAC activity by treatment with the HDACis Vorinostat and Entinostat or Cre-mediated deletion of Hdac1. Changes in overall protein expression as well as histone and protein acetylation were measured following Hdac1 deletion or pharmacological inhibition using label-free liquid chromatography mass spectrometry (LC-MS/MS). We found changes in overall protein abundance and increased acetylation of histones and non-histone proteins, many of which were newly discovered and associated with major metabolic and DNA damage pathways. For non-histone acetylation, we mapped a total of 1204 acetylated peptides corresponding to 603 proteins, including chromatin modifying proteins and transcription factors. Hyperacetylated proteins were involved in processes such as transcription, RNA metabolism and DNA damage repair (DDR). The DDR pathway was majorly affected by hyperacetylation following HDAC inhibition. This included acetylation of H2AX, PARP1 and previously unrecognized acetylation sites in TP53BP1. Our data provide a comprehensive view of the targets of HDAC inhibition in malignant T cells with general applicability and could have translational impact for the treatment of ALCL with HDACis alone or in combination therapies.

• Klíčová slova
• ALCL, MS-275, SAHA, acetylomics, anaplastic large cell lymphoma, entinostat, histone deacetylase inhibitors, histone deacetylases, proteomics, vorinostat,
• MeSH
• acetylace MeSH
• anaplastický velkobuněčný lymfom * farmakoterapie   MeSH
• chromatografie kapalinová MeSH
• histondeacetylasy * metabolismus   MeSH
• histony metabolismus   MeSH
• kyseliny hydroxamové farmakologie   MeSH
• myši MeSH
• tandemová hmotnostní spektrometrie MeSH
• zvířata MeSH
• Check Tag
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• histondeacetylasy * MeSH
• histony MeSH
• kyseliny hydroxamové MeSH

Changes in histone modifications are an attractive model through which environmental signals, such as diet, could be integrated in the cell for regulating its lifespan. However, evidence linking dietary interventions with specific alterations in histone modifications that subsequently affect lifespan remains elusive. We show here that deletion of histone N-alpha-terminal acetyltransferase Nat4 and loss of its associated H4 N-terminal acetylation (N-acH4) extend yeast replicative lifespan. Notably, nat4Δ-induced longevity is epistatic to the effects of calorie restriction (CR). Consistent with this, (i) Nat4 expression is downregulated and the levels of N-acH4 within chromatin are reduced upon CR, (ii) constitutive expression of Nat4 and maintenance of N-acH4 levels reduces the extension of lifespan mediated by CR, and (iii) transcriptome analysis indicates that nat4Δ largely mimics the effects of CR, especially in the induction of stress-response genes. We further show that nicotinamidase Pnc1, which is typically upregulated under CR, is required for nat4Δ-mediated longevity. Collectively, these findings establish histone N-acH4 as a regulator of cellular lifespan that links CR to increased stress resistance and longevity.

• Klíčová slova
• Nat4, Pnc1, calorie restriction, histone N‐terminal acetylation, lifespan,
• MeSH
• acetylace MeSH
• aktivace transkripce MeSH
• časové faktory MeSH
• chromatin metabolismus   MeSH
• dlouhověkost MeSH
• down regulace MeSH
• histonacetyltransferasy metabolismus   MeSH
• histony metabolismus   MeSH
• kalorická restrikce * MeSH
• N-terminální acetyltransferasa D nedostatek   genetika   fyziologie   MeSH
• nikotinamidasa genetika   metabolismus   MeSH
• posttranslační úpravy proteinů MeSH
• regulace genové exprese u hub * MeSH
• Saccharomyces cerevisiae - proteiny genetika   metabolismus   fyziologie   MeSH
• Saccharomyces cerevisiae genetika   fyziologie   MeSH
• stanovení celkové genové exprese MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• chromatin MeSH
• histonacetyltransferasy MeSH
• histony MeSH
• N-terminální acetyltransferasa D MeSH
• NAT4 protein, S cerevisiae MeSH Prohlížeč
• nikotinamidasa MeSH
• PNC1 protein, S cerevisiae MeSH Prohlížeč
• Saccharomyces cerevisiae - proteiny MeSH

Post-translational modification of histones is fundamental to the regulation of basic nuclear processes and subsequent cellular events, including differentiation. In this study, we analyzed acetylated forms of histones H2A, H2B, and H4 during induced differentiation in mouse (mESCs) and human (hESCs) embryonic stem cells and during induced enterocytic differentiation of colon cancer cells in vitro. Endoderm-like differentiation of mESCs induced by retinoic acid and enterocytic differentiation induced by histone deacetylase inhibitor sodium butyrate were accompanied by increased mono-, di-, and tri-acetylation of histone H2B and a pronounced increase in di- and tri-acetylation of histone H4. In enterocytes, mono-acetylation of histone H2A also increased and tetra-acetylation of histone H4 appeared only after induction of this differentiation pathway. During differentiation of hESCs, we observed increased mono-acetylation and decreased tri-acetylation of H2B. Mono-, di-, and tri-acetylation of H4 were reduced, manifested by a significant increase in nonacetylated H4 histones. Levels of acetylated histones increased during induced differentiation in mESCs and during histone deacetylase (HDAC) inhibitor-induced enterocytic differentiation, whereas differentiation of human ESCs was associated with reduced acetylation of histones H2B and H4.

• MeSH
• acetylace MeSH
• buněčná diferenciace MeSH
• embryonální kmenové buňky cytologie   metabolismus   MeSH
• endoderm cytologie   metabolismus   MeSH
• enterocyty cytologie   metabolismus   MeSH
• epigeneze genetická MeSH
• histonacetyltransferasy metabolismus   MeSH
• histony metabolismus   MeSH
• lidé MeSH
• myši MeSH
• nádorové buněčné linie MeSH
• posttranslační úpravy proteinů * MeSH
• spektrometrie hmotnostní - ionizace laserem za účasti matrice MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• histonacetyltransferasy MeSH
• histony MeSH

To study 3D nuclear distributions of epigenetic histone modifications such as H3(K9) acetylation, H3(K4) dimethylation, H3(K9) dimethylation, and H3(K27) trimethylation, and of histone methyltransferase Suv39H1, we used advanced image analysis methods, combined with Nipkow disk confocal microscopy. Total fluorescence intensity and distributions of fluorescently labelled proteins were analyzed in formaldehyde-fixed interphase nuclei. Our data showed reduced fluorescent signals of H3(K9) acetylation and H3(K4) dimethylation (di-me) at the nuclear periphery, while di-meH3(K9) was also abundant in chromatin regions closely associated with the nuclear envelope. Little overlapping (intermingling) was observed for di-meH3(K4) and H3(K27) trimethylation (tri-me), and for di-meH3(K9) and Suv39H1. The histone modifications studied were absent in the nucleolar compartment with the exception of H3(K9) dimethylation that was closely associated with perinucleolar regions which are formed by centromeres of acrocentric chromosomes. Using immunocytochemistry, no di-meH3(K4) but only dense di-meH3(K9) was found for the human acrocentric chromosomes 14 and 22. The active X chromosome was observed to be partially acetylated, while the inactive X was more condensed, located in a very peripheral part of the interphase nuclei, and lacked H3(K9) acetylation. Our results confirmed specific interphase patterns of histone modifications within the interphase nuclei as well as within their chromosome territories.

• MeSH
• acetylace MeSH
• algoritmy MeSH
• buněčné jádro metabolismus   MeSH
• centromera ultrastruktura   MeSH
• fibroblasty metabolismus   MeSH
• histony metabolismus   MeSH
• hybridizace in situ fluorescenční MeSH
• imunohistochemie MeSH
• interfáze fyziologie   MeSH
• lidé MeSH
• lidské chromozomy X genetika   ultrastruktura   MeSH
• lidské chromozomy, pár 14 genetika   ultrastruktura   MeSH
• lidské chromozomy, pár 22 genetika   ultrastruktura   MeSH
• metylace MeSH
• počítačové zpracování obrazu MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• histony MeSH

Molecular changes associated with malignancy are extremely complex. Early epigenetic events occurring in the common tumor types such as breast or prostate cancer might determine the subsequent genetic changes leading to tumor development and progression. Covalent modifications of histones play a major role as determiners of epigenetic information and are important in the regulation of gene expression. Acetylation generally correlates with transcriptional activation, while methylation can signal either activation or repression. However, little is known about the interplay of different epigenetic events. Steroid hormones regulate many cellular processes through signal transduction pathways that result in a variety of post-translational modifications. Such modifications can be triggered by steroid hormones in cooperation with coactivators(p160 family proteins, CBP, p300, p/CAF) and/or corepressors (N-Cor, SMRT, TZF). There is still much to learn about their regulation and the molecular and physiological consequences of these modifications.

• MeSH
• acetylace MeSH
• histony metabolismus   MeSH
• lidé MeSH
• metylace MeSH
• pohlavní steroidní hormony metabolismus   MeSH
• signální transdukce fyziologie   MeSH
• steroidní receptory metabolismus   fyziologie   MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• mužské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• přehledy MeSH
• Názvy látek
• histony MeSH
• pohlavní steroidní hormony MeSH
• steroidní receptory MeSH

Using specific polyclonal antisera raised against acetylated isoforms of histone H4, we have analyzed their distribution in the dioecious plant Silene latifolia (syn. Melandrium album) possessing heteromorphic sex chromosomes. Our previous studies on this species have shown that one of the two X chromosomes in homogametic female cells is heavily methylated and late replicating, as a possible consequence of dosage compensation. Here we report that there are no detectable differences in intensity and distribution of H4 acetylation between these two X chromosomes. In S. latifolia only distal-subtelomeric chromosome regions, on both the sex chromosomes and autosomes, display strong signals of H4 acetylation at N-terminal lysines 5, 8, and 12. These acetylated domains correspond to the very early replicating distal chromosome regions as revealed by 5-bromodeoxyuridine pulses followed by the indirect immunofluorescence microscopy. The distribution of H4 acetylated at lysine 16 was uniform along the chromosomes. The unique distal-subtelomeric H4 acetylation signals were also observed in three other Silene species (S. vulgaris, S. pendula, and S. chalcedonica), but not in two non-related plant species tested (Allium cepa and Nicotiana tobacum). The presented data as well as our recent studies on the structure of S. latifolia chromosome ends indicate that Silene species possess the specific distal-subtelomeric location of euchromatin, gene-rich regions on chromosomes.

• MeSH
• acetylace MeSH
• DNA rostlinná biosyntéza   MeSH
• histony genetika   metabolismus   MeSH
• kinetika MeSH
• králíci MeSH
• replikace DNA * MeSH
• zvířata MeSH
• Check Tag
• králíci MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• DNA rostlinná MeSH
• histony MeSH

OBJECTIVE: Histones bind in a sequence-independent manner to form chromatin. The aminoterminal tails of histones are targets for both phosphorylation and acetylation events. These modifications are thought to fundamentally regulate chromatin structure to accommodate transcription, DNA replication, mitosis and DNA repair. Regeneration of squamous epithelium is accompanied by marked cellular atypia, nuclear and nucleolar pleomorphism which could be confused with neoplasia. The aim of the study was to detect phosphorylated and acetylated forms of histone H3 in cytological smears. DESIGN: Translational research. SETTING: Department of Experimental Oncology, Masaryk Memorial Cancer Institute, Department of Pathological Physiology, Medical Faculty and Department of Obstetrics and Gynecology, Masaryk University, Czech Republic. METHODS: Smears from women aged between 20 to 46 years were selected. The specimens comprised 10 squamous metaplasia, 20 CIN I, 12 CIN II, and 14 CIN III. The smears were stained with polyclonal antibodies against phosphorylated and acetylated forms of histone H3. RESULTS: We found that nuclear positivity for phosphorylated (P) and acetylated (A) forms of histone H3 in CIN II (23% P, 33% A) and CIN III (25% P, 44% A) was higher in comparison with CIN I (8% P, 15% A) and metaplasia (11% P, 12% A). CONCLUSION: We revealed a marked association of histone H3 modifications with the progression of CIN II, CIN III in comparison with CIN I and metaplasia. Our results are in agreement with recent findings: 1. staining of cells with anti-phospho-histone H3 antibodies therefore provides a highly specific marker for mitosis. 2. acetylation of nucleosomal histones correlates with localised transcriptional activity.

• MeSH
• acetylace MeSH
• buněčné jádro metabolismus   MeSH
• cervix uteri metabolismus   patologie   MeSH
• dospělí MeSH
• dysplazie děložního hrdla diagnóza   metabolismus   patologie   MeSH
• fosforylace MeSH
• histony metabolismus   MeSH
• imunohistochemie MeSH
• lidé středního věku MeSH
• lidé MeSH
• metaplazie MeSH
• nádorové biomarkery analýza   MeSH
• nádory děložního čípku diagnóza   metabolismus   patologie   MeSH
• posttranslační úpravy proteinů MeSH
• progrese nemoci MeSH
• vaginální stěr MeSH
• Check Tag
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• histony MeSH
• nádorové biomarkery MeSH