RESUMO
Date palm (Phoenix dactylifera L.) is a dioecious plant, with male and female plants having distinct characteristics. Female plants are responsible for fruit production, and only approximately 10% of male plants are necessary for effective pollination. The determination of plant sex occurs during the first flowering, a process that typically spans 3-7 years. However, this extended timeframe results in significant time and valuable plantation resources being expended in the maintenance of trees. To address this issue, the study focused on sex identification of date palms using DNA markers. The research aimed to develop sex-specific markers for certain date palm cultivars, employing the high annealing temperature random amplified polymorphic DNA (HAT-RAPD) technique for accurate and reliable sex identification. In this investigation, 45 RAPD primers underwent screening in both male and female date palm plants to pinpoint sex-specific markers. Out of the total primers tested, only one, OPW-18, exhibited a correlation with sex. OPW-18 produced a distinct band of approximately 400 bp, consistently present in all male plants but absent in all female plants. The male-specific fragment from OPW-18 was cloned and sequenced to facilitate the development of sex-specific sequence-characterized amplified region (SCAR) primers. The outcomes revealed that the newly crafted SCAR primer pair, mspW18-2F and mspW18-2R, successfully amplified a unique fragment of 283 bp exclusively in male plants. This capability allowed the identification of 100% of male plants in the KL1 and Barhi cultivars. These markers prove to be efficient, reliable, and reproducible for early-stage sex identification in plants.
RESUMO
DNA transposons are diverse in fish genomes and have been described to generate genomic evolutionary novelties. hAT transposable element data are scarce in Teleostei genomes, making it challenging to conduct comparative genomic studies to understand their neutrality or function. This study aimed to perform a genomic and molecular characterization of hAT copies to assess the diversity of these elements and associate changes in these sequences to genomic and karyotypic novelties in Apareiodon sp. The data revealed that hAT TEs are highly abundant in the Apareiodon sp. genome, with few possibly autonomous copies. Highly conserved sequences with likely functional transposases were observed in nine hAT elements. A great diversity of hAT subgroups was observed, especially from Ac, Charlie, Blackjack, Tip100, hAT6, and hAT5, and a similar wave of hAT genomic invasion was identified in the genome for these six groups of hAT sequences. The data also revealed a distinct number of microsatellites within degenerated hAT copies. hAT sites were demonstrated to be dispersed in the Apareiodon sp. chromosomes and not involved in W chromosome-specific region differentiation. In conclusion, the genomic analysis revealed a great diversity of hAT elements, possible autonomous copies, and differentiation of degenerated transposable elements into tandem sequences.
Assuntos
Elementos de DNA Transponíveis , Genoma , Filogenia , Elementos de DNA Transponíveis/genética , Animais , Genoma/genética , Evolução Molecular , Repetições de Microssatélites/genética , Genômica/métodos , Peixes/genética , Peixes/classificaçãoRESUMO
This article describes the diffraction pattern (2-periodic Fourier transform) from the vertices of a large patch of the recently discovered `Spectre' tiling - a strictly chiral aperiodic monotile. It was reported recently that the diffraction pattern of the related weakly chiral aperiodic `Hat' monotile was 2-periodic with chiral plane-group symmetry p6 [Kaplan et al. (2024). Acta Cryst. A80, 72-78]. The diffraction periodicity arises because the Hat tiling is a systematic aperiodic deletion of vertices from the 2-periodic hexagonal mta tiling. Despite the similarity of the Hat and Spectre tilings, the Spectre tiling is not aligned with a 2-periodic lattice, and its diffraction pattern is non-periodic with chiral point symmetry 6 about the origin.
RESUMO
Brassica rapa L. is an important overwintering oilseed crop in Northwest China. Histone acetyltransferases (HATs) play an important role in epigenetic regulation, as well as the regulation of plant growth, development, and responses to abiotic stresses. To clarify the role of histone acetylation in the low-temperature response of B. rapa L., we identified 29 HAT genes in B. rapa L. using bioinformatics tools. We also conducted a comprehensive analysis of the physicochemical properties, gene structure, chromosomal localization, conserved structural domains and motifs, cis-acting regulatory elements, and evolutionary relationships of these genes. Using transcriptome data, we analyzed the expression patterns of BrHAT family members and predicted interactions between proteins; the results indicated that BrHATs play an important role in the low-temperature response of B. rapa L. HAT inhibitor (curcumin; CUR) and histone deacetylase inhibitor (Trichostatin A; TSA) were applied to four B. rapa L. varieties varying in cold resistance under the same low-temperature conditions, and changes in the physiological indexes of these four varieties were analyzed. The inhibitor treatment attenuated the effect of low temperature on seed germination, and curcumin treatment was most effective, indicating that the germination period was primarily regulated by histone acetylase. Both inhibitor treatments increased the activity of protective enzymes and the content of osmoregulatory substances in plants, suggesting that histone acetylation and deacetylation play a significant role in the response of B. rapa L. to low-temperature stress. The qRT-PCR analyses showed that the expression patterns of BrHATs were altered under different inhibitor treatments and low-temperature stress; meanwhile, we found three significantly differentially expressed genes. In sum, the process of histone acetylation is involved in the cold response and the BrHATs gene plays a role in the cold stress response.
Assuntos
Brassica rapa , Temperatura Baixa , Regulação da Expressão Gênica de Plantas , Histona Acetiltransferases , Inibidores de Histona Desacetilases , Histona Acetiltransferases/metabolismo , Histona Acetiltransferases/genética , Brassica rapa/genética , Brassica rapa/efeitos dos fármacos , Brassica rapa/crescimento & desenvolvimento , Brassica rapa/metabolismo , Regulação da Expressão Gênica de Plantas/efeitos dos fármacos , Inibidores de Histona Desacetilases/farmacologia , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Família Multigênica , Germinação/efeitos dos fármacos , Filogenia , Acetilação/efeitos dos fármacosRESUMO
BACKGROUND: Surveillance of "silent" human African Trypanosomiasis (HAT) foci is important for the achievement of the World Health Organization (WHO) goal of interrupting the transmission of this disease by 2030. It is in this context that this study was carried out to determine the trypanosome species circulating in the "silent" HAT foci of Bafia and the Manoka island in Cameroon. METHODS: In the Bafia and Manoka HAT foci, georeferenced pyramidal traps were used to trap tsetse flies. After DNA extraction from each whole fly, molecular tools were used to detect different trypanosome species as well as the origin of tsetse fly blood meals. Geographical information system was used to map the trypanosome infections and entomological data and to localize areas at high risk for trypanosome transmission. RESULTS: For this study, 1683 tsetse flies were caught and the relative apparent densities was 2.96: 0.03 in the Bafia HAT focus and 5.23 in the Manoka island. For the molecular identification of trypanosomes, 708 non-teneral tsetse flies (8 from Bafia and 700 from Manoka) were randomly selected. The overall trypanosome infection rate was 7.34 % with no infection in the Bafia HAT focus. Among the analysed flies, 4.57 % had trypanosomes of the subgenus Trypanozoon while 4.1 % and 1.13 % had respectively T. congolense and T. vivax. The most common mixed infections were the combination of trypanosomes of the subgenus Trypanozoon and T. congolense. Of the 708 tsetse flies analysed, 134 (18.93 %) tsetse flies were found with residual blood meals, 94 % and 6 % were respectively from humans and dogs. The trapping sites of Plateau, Sandje and Hospital appeared as the areas where contact with tsetse flies is most common. CONCLUSION: This study revealed a discrepancy in the abundance tsetse flies as well as the trypanosome infection rates in tsetse of the two "silent" HAT foci of Cameroon. The detection of different trypanosome species in tsetse from the Manoka Island highlights their transmission. The high percentage of human blood meals in tsetse flies indicates an important contact between tsetse flies and human; emphasizing the risk of trypanosome transmission to human in this island.
RESUMO
MAIN CONCLUSION: This review focuses on HATs and HDACs that modify non-histone proteins, summarizes functional mechanisms of non-histone acetylation as well as the roles of HATs and HDACs in rice and Arabidopsis. The growth and development of plants, as well as their responses to biotic and abiotic stresses, are governed by intricate gene and protein regulatory networks, in which epigenetic modifying enzymes play a crucial role. Histone lysine acetylation levels, modulated by histone acetyltransferases (HATs) and histone deacetylases (HDACs), are well-studied in the realm of transcriptional regulation. However, the advent of advanced proteomics has unveiled that non-histone proteins also undergo acetylation, with its underlying mechanisms now being clarified. Indeed, non-histone acetylation influences protein functionality through diverse pathways, such as modulating protein stability, adjusting enzymatic activity, steering subcellular localization, influencing interactions with other post-translational modifications, and managing protein-protein and protein-DNA interactions. This review delves into the recent insights into the functional mechanisms of non-histone acetylation in plants. We also provide a summary of the roles of HATs and HDACs in rice and Arabidopsis, and explore their potential involvement in the regulation of non-histone proteins.
Assuntos
Arabidopsis , Histona Acetiltransferases , Histona Desacetilases , Oryza , Proteínas de Plantas , Processamento de Proteína Pós-Traducional , Histona Desacetilases/metabolismo , Histona Desacetilases/genética , Acetilação , Oryza/genética , Oryza/metabolismo , Oryza/enzimologia , Arabidopsis/genética , Arabidopsis/metabolismo , Arabidopsis/enzimologia , Histona Acetiltransferases/metabolismo , Histona Acetiltransferases/genética , Proteínas de Plantas/metabolismo , Proteínas de Plantas/genética , Regulação da Expressão Gênica de Plantas , Histonas/metabolismoRESUMO
In order to study the influence of modular filled and composite material forms on the axial crushing and energy absorption properties of structures, modular filled composite structures were constructed, and innovatively applied to the inner side of double-hat beam (DHB) structures in automobiles. The modular filled structures comprise hexagonal, quadrilateral, and triangular sections. By analyzing the collision performance of modular filled DHB structures, significant enhancements were observed in both the sectional characteristics and the specific Mean Crushing Force of modular filled DHBs compared to the conventional double-hat beam structure. These advancements notably improved the plastic deformation characteristics of the structures. Additionally, dynamic weightlessness experiments were conducted to validate the accuracy of the simulation model. Among the proposed schemes, namely QU-5, HE-5, and TR-5, notable improvements in crashworthiness were identified. Specifically, crashworthiness indicators increased by 32.54%, 78.9%, and 116.53%. Compared with other thin-walled structures, modular filled composite DHBs have advantages in axial crushing and energy absorption. By optimizing layout characteristics, the modular filled structures will achieve significant lightweight and energy absorption performance improvements. This work has clear reference value for automotive engineers and scholars to further explore the axial crash safety, platform modularization, and lightweight design of vehicles.
RESUMO
The control of African trypanosomiasis (AT) in Eastern and Southern Africa, including Zambia, faces huge challenges due to the involvement of wild and domestic animal reservoirs. Free-roaming dogs in wildlife-populated and tsetse-infested villages of Zambia's Mambwe district are exposed to infectious tsetse bites. Consuming fresh raw game meat and bones further exacerbates their risk of contracting AT. We focus on the reservoir role of such dogs in maintaining and transmitting diverse species of trypanosomes that are infective to humans and livestock in Zambia's Mambwe district. A cohort of 162 dogs was enrolled for follow-up at 3 different time points from June to December 2018 in selected villages of Malama, Mnkhanya, and Nsefu chiefdoms of Mambwe district, eastern Zambia. Blood and serum were screened for AT by microscopy, GM6 ELISA, PCR (ITS1 and SRA), and Sanger sequencing. Out of the 162 dogs in the cohort, 40 were lost to follow-up and only 122 remained traceable at the end of the study. GM6 ELISA detected Trypanosoma antibodies in 121 dogs (74.7%) and ITS1-PCR detected DNA involving single and mixed infections of T. congolense, T. brucei, and suspected T. simiae or T. godfreyi in 115 dogs (70.9%). The human-infective T. b. rhodesiense was detected by SRA PCR in 67 dogs (41.4%), and some sequence data that support the findings of this study have been deposited in the GenBank under accession numbers OL961811, OL961812, and OL961813. Our study demonstrates that the Trypanosoma reservoir community in Zambia is wider than was thought and includes domesticated dogs. As dogs are active carriers of human and livestock-infective trypanosomes, they pose a risk of transmitting AT in endemic villages of Mambwe district as they are neglected and left untreated. To fully bring AT under control, countries such as Zambia where the role of animal reservoirs is important, should not limit their prevention and treatment efforts to livestock (especially cattle) but also include dogs that play an integral part in most rural communities.
Assuntos
Reservatórios de Doenças , Doenças do Cão , Tripanossomíase Africana , Animais , Cães , Zâmbia/epidemiologia , Tripanossomíase Africana/epidemiologia , Tripanossomíase Africana/veterinária , Tripanossomíase Africana/transmissão , Tripanossomíase Africana/parasitologia , Doenças do Cão/parasitologia , Doenças do Cão/epidemiologia , Doenças do Cão/transmissão , Reservatórios de Doenças/parasitologia , Humanos , Masculino , Feminino , Animais Domésticos/parasitologia , Anticorpos Antiprotozoários/sangue , Trypanosoma/genética , Trypanosoma/isolamento & purificaçãoRESUMO
Histone acetyltransferase inhibitors (HATi) are mechanism-based inhibitors that show promise in the treatment of several illnesses, including diabetes, hyperlipidemia, cancer, and Alzheimer's disease. The work emphasizes the significance of HATi as a possible treatment strategy against Candida species biofilms. Here, in this study, we found that combining a HATi, anacardic acid (AA), and quercetin, a known flavonoid, significantly prevented biofilm formation by C. tropicalis. We further show that C. tropicalis exhibited a considerable downregulation of drug-resistance gene expression (CDR1 and MDR1) when co-administrated. Additionally, in silico studies revealed that the AA interacts strongly with a histone acetyltransferase, Rtt109, which may account for the observed biofilm inhibitory effect. In conclusion, the study illustrates how HATi may be used to potentiate the inhibitory action of phytoactives or antifungals against drug-resistant yeast infections.
Assuntos
Ácidos Anacárdicos , Antifúngicos , Biofilmes , Candida tropicalis , Sinergismo Farmacológico , Histona Acetiltransferases , Quercetina , Candida tropicalis/efeitos dos fármacos , Quercetina/farmacologia , Biofilmes/efeitos dos fármacos , Antifúngicos/farmacologia , Histona Acetiltransferases/antagonistas & inibidores , Histona Acetiltransferases/metabolismo , Histona Acetiltransferases/genética , Ácidos Anacárdicos/farmacologia , Farmacorresistência Fúngica , Testes de Sensibilidade Microbiana , Inibidores Enzimáticos/farmacologia , Proteínas Fúngicas/genética , Proteínas Fúngicas/metabolismo , Proteínas Fúngicas/antagonistas & inibidoresRESUMO
Infrared matrix-assisted laser desorption electrospray ionization (IR-MALDESI) uses an infrared laser to desorb neutral biomolecules with postionization via ESI at atmospheric pressure. The Gaussian profile of the laser with conventional optics results in the heating of adjacent nonablated tissue due to the energy profile being circular. A diffractive optical element (DOE) was incorporated into the optical train to correct for this disadvantage. The DOE produces a top-hat beam profile and square ablation spots, which have uniform energy distributions. Although beneficial to mass spectrometry imaging (MSI), it is unknown how the DOE affects the ability to perform quantitative MSI (qMSI). In this work, we evaluate the performance of the DOE optical train against our conventional optics to define the potential advantages of the top-hat beam profile. Absolute quantification of glutathione (GSH) was achieved by normalizing the analyte of interest to homoglutathione (hGSH), spotting a dilution series of stable isotope labeled glutathione (SIL-GSH), and analyzing by IR-MALDESI MSI with either the conventional optical train or with the DOE incorporated. Statistical comparison indicates that there was no significant difference between the quantification of GSH by the two optical trains as evidenced by similar calibration curves. Results support that both optical trains can be used for qMSI without a change in the ability to carry out absolute quantification but providing the benefits of the top-hat optical train (i.e., flat energy profile and square ablation spots)-for future qMSI studies.
Assuntos
Glutationa , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz , Glutationa/análise , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz/métodos , AnimaisRESUMO
Photocatalytic reduction reactions occasionally utilize sacrificial agents to scavenge photogenerated holes, thus enhancing the kinetics and efficiency of electron harvesting. However, exploring alternative hole-mediated oxidation reactions and their potential impact on photoredox processes is limited. This study investigates the products resulting from the oxidation of ethanol, a commonly used hole scavenger, and the underlying mechanisms involved. We examine a homogeneous eosin Y photoreaction scheme containing a Cu complex coordinated with an N-heterocyclic carbene, a combination often employed in CO2 conversion. Under visible-light excitation, this photosystem yields methane as an unusual product, alongside acetaldehyde and carbon monoxide. Mechanistic analysis reveals that ethanol undergoes a catalytic cascade involving oxidative processes, C-C bond cleavage, and intermolecular hydrogen atom transfer. Notably, the Lewis-acidic metal center of the Cu complex activates a novel pathway for ethanol oxidation. This work presents the influence of catalyst selection and reaction condition optimization on the emergence of new or unexpected catalytic processes.
RESUMO
Excessive fluoride ingestion during tooth development can cause dental fluorosis. Previously, we reported that fluoride activates histone acetyltransferase (HAT) to acetylate p53, promoting fluoride toxicity in mouse ameloblast-like LS8 cells. However, the roles of HAT and histone acetylation status in fluoride-mediated gene expression remain unidentified. Here, we demonstrate that fluoride-mediated histone modification causes gene expression alterations in LS8 cells. LS8 cells were treated with or without fluoride followed by ChIP-Seq analysis of H3K27ac. Genes were identified by differential H3K27ac peaks within ±1 kb from transcription start sites. The levels of mRNA of identified genes were assessed using rea-time PCR (qPCR). Fluoride increased H3K27ac peaks associated with Bax, p21, and Mdm2 genes and upregulated their mRNA levels. Fluoride decreased H3K27ac peaks and p53, Bad, and Bcl2 had suppressed transcription. HAT inhibitors (Anacardic acid or MG149) suppressed fluoride-induced mRNA of p21 and Mdm2, while fluoride and the histone deacetylase (HDAC) inhibitor sodium butyrate increased Bad and Bcl2 expression above that of fluoride treatment alone. To our knowledge, this is the first study that demonstrates epigenetic regulation via fluoride treatment via H3 acetylation. Further investigation is required to elucidate epigenetic mechanisms of fluoride toxicity in enamel development.
Assuntos
Ameloblastos , Fluoretos , Histonas , Animais , Camundongos , Acetilação/efeitos dos fármacos , Histonas/metabolismo , Ameloblastos/metabolismo , Ameloblastos/efeitos dos fármacos , Fluoretos/farmacologia , Fluoretos/toxicidade , Linhagem Celular , Regulação da Expressão Gênica/efeitos dos fármacos , Histona Acetiltransferases/metabolismo , Histona Acetiltransferases/genética , Epigênese Genética/efeitos dos fármacos , Inibidores de Histona Desacetilases/farmacologiaRESUMO
The external appearance of fruit commodities is an essential trait that has profound effects on consumer preferences. A natural melon variety, characterized by an uneven and patchy arrangement of dark green streaks and spots on the white-skinned rind, resembles shooting stars streaking across the sky; thus, this variety is called "Shooting Star" (SS). To investigate the mechanism underlying the SS melon rind pattern, we initially discovered that the variegated dark green color results from chlorophyll accumulation on the white skin. We then constructed a segregation population by crossing a SS inbred line with a white rind (WR) inbred line and used bulk segregant analysis (BSA) revealed that the SS phenotype is controlled by a single dominant gene, CmAPRR2, which has been previously confirmed to determine dark green coloration. Further genomic analysis revealed a hAT-like transposable element (TE) inserted in CmAPRR2. This TE in CmAPRR2 is recurrently excised from rind tissues, activating the expression of CmAPRR2. This activation promotes the accumulation of chlorophyll, leading to the variegated dark green color on the rind, and ultimately resulting in the SS rind phenotype. Therefore, we propose that the SS phenotype results from the recurrent excision of the hAT-like TE in CmAPRR2.
Assuntos
Elementos de DNA Transponíveis , Fenótipo , Elementos de DNA Transponíveis/genética , Pigmentação/genética , Frutas/genética , Clorofila/metabolismo , Regulação da Expressão Gênica de Plantas , Cucurbitaceae/genética , Cucurbitaceae/metabolismo , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Genes de Plantas/genéticaRESUMO
Glioblastoma (GBM) is a fatal adult brain tumor with an extremely poor prognosis. GBM poses significant challenges for targeted therapies due to its intra- and inter-tumoral heterogeneity, a highly immunosuppressive microenvironment, diffuse infiltration into normal brain parenchyma, protection by the blood-brain barrier and acquisition of therapeutic resistance. Recent studies have implicated epigenetic modifiers as key players driving tumorigenesis, resistance, and progression of GBM. While the vast majority of GBM research on epigenetic modifiers thus far has focused predominantly on elucidating the functional roles and targeting of DNA methyltransferases and histone deacetylases, emerging evidence indicates that histone acetyltransferases (HATs) also play a key role in mediating plasticity and therapeutic resistance in GBM. Here, we will provide an overview of HATs, their dual roles and functions in cancer as both tumor suppressors and oncogenes and focus specifically on their implications in GBM resistance. We also discuss the technical challenges in developing selective HAT inhibitors and highlight their promise as potential anti-cancer therapeutics for treating intractable cancers such as GBM.
Assuntos
Neoplasias Encefálicas , Resistencia a Medicamentos Antineoplásicos , Glioblastoma , Histona Acetiltransferases , Humanos , Glioblastoma/tratamento farmacológico , Glioblastoma/genética , Glioblastoma/enzimologia , Glioblastoma/patologia , Histona Acetiltransferases/antagonistas & inibidores , Histona Acetiltransferases/metabolismo , Histona Acetiltransferases/genética , Neoplasias Encefálicas/tratamento farmacológico , Neoplasias Encefálicas/genética , Neoplasias Encefálicas/enzimologia , Neoplasias Encefálicas/patologia , Epigênese Genética , Terapia de Alvo Molecular , Animais , Antineoplásicos/uso terapêutico , Antineoplásicos/farmacologia , Regulação Neoplásica da Expressão Gênica/efeitos dos fármacosRESUMO
Gaseous alkanes represent the most abundant carbon-based chemical feedstocks in our planet. However, the intrinsic inertness of their C-H bonds has rendered the use of these alkanes very difficult for purposes beyond aerobic combustion and energy intensive processes. Thus, clean and energy-efficient transformations for their use in synthetic organic chemistry are still rare. Here we report a catalytic methodology for the direct cross-coupling of gaseous alkanes with (hetero)aryl bromides through the combination of metallaphotoredox-mediated hydrogen atom transfer and nickel catalysis. This protocol provides an efficient platform for the addition of short alkyl groups into diverse (hetero) aromatic rings, providing a wide range of high-value alkyl(hetero)arenes, and bypassing the longstanding need of using preactivated alkylating agents in C(sp2)-C(sp3) cross-couplings. The method features high chemoselectivity, regioselectivity and a remarkable functional group tolerance, operates under mild conditions, and exhibits operational simplicity.
RESUMO
Nucleosides with a substituent at the 4'-position have received much attention as antiviral drugs and as raw materials for oligonucleotide therapeutics. 4'-Modified nucleosides are generally synthesized using ionic reactions through the introduction of electrophilic or nucleophilic substituents at the 4'-position. However, their synthetic methods have some drawbacks; e.g., (i) it is difficult to control stereoselectivity at the 4'-position; (ii) complex protection-deprotection processes are required; (iii) the range of electrophiles and nucleophiles is limited. With this background, we considered that a carbon radical generated at the 4'-position would be a useful intermediate for the synthesis of 4'-modified nucleosides. In this review, two novel methods for the generation of 4'-carbon radicals are summarized. The first utilizes radical deformylation involving ß-fragmentation of a hydroxymethyl group at the 4'-position. The other utilizes radical decarboxylation and 1,5-hydrogen atom transfer (1,5-HAT), which enables the generation of 4'-carbon radicals while retaining the hydroxymethyl group at the 4'-position. These methods enable the rapid and facile generation of 4'-carbon radicals and provide various 4'-modified nucleosides including 2',4'-bridged structures.
Assuntos
Antivirais , Carbono , Nucleosídeos , Nucleosídeos/síntese química , Nucleosídeos/química , Carbono/química , Radicais Livres/química , Radicais Livres/síntese química , Antivirais/síntese química , Antivirais/química , Técnicas de Química Sintética/métodos , Hidrogênio/químicaRESUMO
BACKGROUND: Histone deacetylases (HDACs) are crucial regulators of gene expression, DNA synthesis, and cellular processes, making them essential targets in cancer research. HDAC6, specifically, influences protein stability and chromatin dynamics. Despite HDAC6's potential therapeutic value, its exact role in gene regulation and chromatin remodeling needs further clarification. This study examines how HDAC6 inactivation influences lysine acetyltransferase P300 stabilization and subsequent effects on chromatin structure and function in cancer cells. METHODS AND RESULTS: We employed the HDAC6 inhibitor ITF3756, siRNA, or CRISPR/Cas9 gene editing to inactivate HDAC6 in different epigenomic backgrounds. Constantly, this inactivation led to significant changes in chromatin accessibility, particularly increased acetylation of histone H3 lysines 9, 14, and 27 (ATAC-seq and H3K27Ac ChIP-seq analysis). Transcriptomics, proteomics, and gene ontology analysis revealed gene changes in cell proliferation, adhesion, migration, and apoptosis. Significantly, HDAC6 inactivation altered P300 ubiquitination, stabilizing P300 and leading to downregulating genes critical for cancer cell survival. CONCLUSIONS: Our study highlights the substantial impact of HDAC6 inactivation on the chromatin landscape of cancer cells and suggests a role for P300 in contributing to the anticancer effects. The stabilization of P300 with HDAC6 inhibition proposes a potential shift in therapeutic focus from HDAC6 itself to its interaction with P300. This finding opens new avenues for developing targeted cancer therapies, improving our understanding of epigenetic mechanisms in cancer cells.
Assuntos
Cromatina , Desacetilase 6 de Histona , Inibidores de Histona Desacetilases , Humanos , Desacetilase 6 de Histona/genética , Desacetilase 6 de Histona/antagonistas & inibidores , Cromatina/genética , Cromatina/efeitos dos fármacos , Linhagem Celular Tumoral , Inibidores de Histona Desacetilases/farmacologia , Acetilação/efeitos dos fármacos , Neoplasias/tratamento farmacológico , Neoplasias/genética , Proteína p300 Associada a E1A/genética , Proteína p300 Associada a E1A/metabolismo , Regulação Neoplásica da Expressão Gênica/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Proliferação de Células/genética , Histonas/metabolismo , Ubiquitinação/efeitos dos fármacosRESUMO
Primary malignancies of the central nervous system account for 2% of all cancers in adults and almost 15% in children under 15 years of age. The prognosis of brain anaplastic cancers and glioblastomas remains extremely poor, with devastating survival expectative, and new molecular markers and therapeutic targets are essential. Epigenetic changes constitute an extensive field for the development of new diagnostic and therapeutic strategies. Histone acetyl transferase-1 (HAT1) has merged as a potential prognostic marker and therapy target for different malignancies. Data repository analysis showed HAT1 mRNA overexpression in gliomas and has been described its alternative splicing in glioblastomas. Using immunohistochemical and aptahistochemical methods, we analyzed the expression of HAT1 in meningiomas, oligodendrogliomas, and astroglial cancers. We observed that HAT1 overexpression is associated with the most aggressive tumor types and the worse prognosis, as well as with a higher probability of early relapse in meningiomas. Its cytosolic localization correlates with tumor progression and prognosis. Aptamers, synthetic oligonucleotides capable to bind and inhibit a wide variety of targets, are considered as promising diagnostic and therapeutic tools. Aptahistochemistry using the aptamer apHAT610 offered superior results in comparison with the antibody used, as a good example of the potential of aptamers as diagnostic tools for histopathology.
Assuntos
Neoplasias Encefálicas , Histona Acetiltransferases , Imuno-Histoquímica , Neoplasias Meníngeas , Humanos , Neoplasias Encefálicas/patologia , Neoplasias Encefálicas/diagnóstico , Neoplasias Encefálicas/metabolismo , Neoplasias Encefálicas/genética , Neoplasias Meníngeas/patologia , Neoplasias Meníngeas/diagnóstico , Neoplasias Meníngeas/genética , Neoplasias Meníngeas/metabolismo , Prognóstico , Feminino , Masculino , Pessoa de Meia-Idade , Histona Acetiltransferases/genética , Histona Acetiltransferases/metabolismo , Histona Acetiltransferases/análise , Adulto , Idoso , Glioma/patologia , Glioma/diagnóstico , Glioma/metabolismo , Glioma/genética , Meningioma/patologia , Meningioma/diagnóstico , Meningioma/genética , Meningioma/metabolismo , Meningioma/enzimologia , Biomarcadores Tumorais/genética , Biomarcadores Tumorais/metabolismo , Biomarcadores Tumorais/análise , Gradação de TumoresRESUMO
BACKGROUND: The transposons of the hAT superfamily are the most widespread transposons ever known. SLEEPER genes encode domesticated transposases from the hAT superfamily, which may have lost their transposable functions during long-term evolution and transformed into host proteins that regulate plant growth and development. RESULTS: This study identified 162 members of the SLEEPER gene family from Brassica napus. These members are widely distributed on 19 chromosomes, mainly in the Cn subgenome, and have promoters with various cis-acting elements related to hormone regulation, abiotic stress, and growth and development regulation. Most of the genes in this family contain similar conserved domains and motifs, and the closer the genes are distributed on evolutionary branches, the more similar their structures are. Transcriptome sequencing performed on tissues at different growth stages from B. napus line 3529 indicated that these genes had different expression patterns, and nearly half of the genes were not detectably expressed in all samples. CONCLUSIONS: This study investigated the gene structure, expression patterns, evolutionary features, and gene localization of the SLEEPER family members to confirm the significance of these genes in the growth of B. napus, providing a reference for the study of transposon domestication and outstanding genetic resources for the genetic improvement of B. napus.
Assuntos
Brassica napus , Elementos de DNA Transponíveis , Regulação da Expressão Gênica de Plantas , Família Multigênica , Brassica napus/genética , Brassica napus/metabolismo , Elementos de DNA Transponíveis/genética , Genoma de Planta , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Genes de Plantas , Filogenia , Transposases/genética , Transposases/metabolismo , Evolução Molecular , Perfilação da Expressão GênicaRESUMO
Acetylation of histone proteins by histone acetyltransferases (HATs), and the resultant change in gene expression, is a well-established mechanism necessary for long-term memory (LTM) consolidation, which is not required for short-term memory (STM). However, we previously demonstrated that the HAT p300/CBP-associated factor (PCAF) also influences hippocampus (HPC)-dependent STM in male rats. In addition to their epigenetic activity, HATs acetylate nonhistone proteins involved in nongenomic cellular processes, such as estrogen receptors (ERs). Given that ERs have rapid, nongenomic effects on HPC-dependent STM, we investigated the potential interaction between ERs and PCAF for STM mediated by the dorsal hippocampus (dHPC). Using a series of pharmacological agents administered directly into the dHPC, we reveal a functional interaction between PCAF and ERα in the facilitation of short-term object-in-place memory in male but not female rats. This interaction was specific to ERα, while ERß agonism did not enhance STM. It was further specific to dHPC STM, as the effect was not present in the dHPC for LTM or in the perirhinal cortex. Further, while STM required local (i.e., dHPC) estrogen synthesis, the facilitatory interaction effect appeared independent of estrogens. Finally, western blot analyses demonstrated that PCAF activation in the dHPC rapidly (5 min) activated downstream estrogen-related cell signaling kinases (c-Jun N-terminal kinase and extracellular signal-related kinase). Collectively, these findings indicate that PCAF, which is typically implicated in LTM through epigenetic processes, also influences STM in the dHPC, possibly via nongenomic ER activity. Critically, this novel PCAF-ER interaction might exist as a male-specific mechanism supporting STM.