RESUMO
Despite the undisputed development of medicine, antibiotics still serve as first-choice drugs for patients with infectious disorders. The widespread use of antibiotics results from a wide spectrum of their actions encompassing mechanisms responsible for: the inhibition of bacterial cell wall biosynthesis, the disruption of cell membrane integrity, the suppression of nucleic acids and/or proteins synthesis, as well as disturbances of metabolic processes. However, the widespread availability of antibiotics, accompanied by their overprescription, acts as a double-edged sword, since the overuse and/or misuse of antibiotics leads to a growing number of multidrug-resistant microbes. This, in turn, has recently emerged as a global public health challenge facing both clinicians and their patients. In addition to intrinsic resistance, bacteria can acquire resistance to particular antimicrobial agents through the transfer of genetic material conferring resistance. Amongst the most common bacterial resistance strategies are: drug target site changes, increased cell wall permeability to antibiotics, antibiotic inactivation, and efflux pumps. A better understanding of the interplay between the mechanisms of antibiotic actions and bacterial defense strategies against particular antimicrobial agents is crucial for developing new drugs or drug combinations. Herein, we provide a brief overview of the current nanomedicine-based strategies that aim to improve the efficacy of antibiotics.
Assuntos
Anti-Infecciosos , Infecções Bacterianas , Humanos , Antibacterianos/farmacologia , Antibacterianos/uso terapêutico , Antibacterianos/metabolismo , Bactérias/metabolismo , Infecções Bacterianas/tratamento farmacológicoRESUMO
Fucosidosis is a rare neurodegenerative autosomal recessive disorder, which manifests as progressive neurological and psychomotor deterioration, growth retardation, skin and skeletal abnormalities, intellectual disability and coarsening of facial features. It is caused by biallelic mutations in FUCA1 encoding the α-L-fucosidase enzyme, which in turn is responsible for degradation of fucose-containing glycoproteins and glycolipids. FUCA1 mutations lead to severe reduction or even loss of α-L-fucosidase enzyme activity. This results in incomplete breakdown of fucose-containing compounds leading to their deposition in different tissues and, consequently, disease progression. To date, 36 pathogenic variants in FUCA1 associated with fucosidosis have been documented. Among these are three splice site variants. Here, we report a novel fucosidosis-related 9-base-pair deletion (NG_013346.1:g.10233_10241delACAGGTAAG) affecting the exon 3/intron 3 junction within a FUCA1 sequence. This novel pathogenic variant was identified in a five-year-old Polish girl with a well-defined pattern of fucosidosis symptoms. Since it is postulated that other genetic, nongenetic or environmental factors can also contribute to fucosidosis pathogenesis, we performed further analysis and found two rare de novo chromosomal aberrations in the girl's genome involving a 15q11.1-11.2 microdeletion and an Xq22.2 gain. These abnormalities were associated with genome-wide changes in DNA methylation status in the epigenome of blood cells.
Assuntos
Aberrações Cromossômicas , Fucosidose/genética , alfa-L-Fucosidase/genética , Pré-Escolar , Cromossomos Humanos Par 15/genética , Cromossomos Humanos X/genética , Metilação de DNA , Análise Mutacional de DNA , Feminino , Fucosidose/diagnóstico , Humanos , Polônia , Deleção de SequênciaRESUMO
The aim of this study was to examine novel putative markers of the response to the competitive soccer match in adolescent players, such as changes in global levels of γH2AX and H4K16ac in the chromatin of peripheral mononuclear blood cells (PMBCs) and a Fourier-transform infrared spectroscopy (FTIR)-based biochemical fingerprint of serum. These characteristics were examined with reference to the physiological and metabolic aspects of this response. Immediately post-match we noticed: (1) a systemic inflammatory response, manifesting as peaks in leukocyte count and changes in concentrations of IL-6, TNFα, and cortisol; (2) a peak in plasma lactate; (3) onset of oxidative stress, manifesting as a decline in GSH/GSSG; (4) onset of muscle injury, reflected in an increase in CK activity. Twenty-four hours post-match the decrease in GSH/GSSG was accompanied by accumulation of MDA and 8-OHdG, macromolecule oxidation end-products, and an increase in CK activity. No changes in SOD1 or GPX1 levels were found. Repeated measures correlation revealed several associations between the investigated biomarkers. The FTIR analysis revealed that the match had the greatest impact on serum lipid profile immediately post-game. In turn, increases in γH2AX and H4K16ac levels at 24 h post-match indicated activation of a DNA repair pathway.
Assuntos
Desempenho Atlético , Histonas/metabolismo , Futebol/fisiologia , Adolescente , Antropometria , Biomarcadores/sangue , Creatina Quinase/sangue , Reparo do DNA , Feminino , Humanos , Hidrocortisona/sangue , Inflamação , Interleucina-6/sangue , Ácido Láctico/sangue , Leucócitos Mononucleares/citologia , Substâncias Macromoleculares , Masculino , Fadiga Muscular , Músculo Esquelético/fisiologia , Estresse Oxidativo , Espectroscopia de Infravermelho com Transformada de Fourier , Fator de Necrose Tumoral alfa/sangueRESUMO
Monocytes, which play a crucial role in the immune system, are characterized by an enormous sensitivity to oxidative stress. As they lack four key proteins responsible for DNA damage response (DDR) pathways, they are especially prone to reactive oxygen species (ROS) exposure leading to oxidative DNA lesions and, consequently, ROS-driven apoptosis. Although such a phenomenon is of important biological significance in the regulation of monocyte/macrophage/dendritic cells' balance, it also a challenge for monocytic mechanisms that have to provide and maintain genetic stability of its own DNA. Interestingly, apurinic/apyrimidinic endonuclease 1 (APE1), which is one of the key proteins in two DDR mechanisms, base excision repair (BER) and non-homologous end joining (NHEJ) pathways, operates in monocytic cells, although both BER and NHEJ are impaired in these cells. Thus, on the one hand, APE1 endonucleolytic activity leads to enhanced levels of both single- and double-strand DNA breaks (SSDs and DSBs, respectively) in monocytic DNA that remain unrepaired because of the impaired BER and NHEJ. On the other hand, there is some experimental evidence suggesting that APE1 is a crucial player in monocytic genome maintenance and stability through different molecular mechanisms, including induction of cytoprotective and antioxidant genes. Here, the dual face of APE1 is discussed.
Assuntos
DNA Liase (Sítios Apurínicos ou Apirimidínicos)/genética , Sistema Imunitário/metabolismo , Monócitos/metabolismo , Estresse Oxidativo/genética , Reparo do DNA por Junção de Extremidades/genética , Reparo do DNA/genética , Células Dendríticas/imunologia , Células Dendríticas/metabolismo , Humanos , Macrófagos/imunologia , Macrófagos/metabolismo , Monócitos/imunologia , Espécies Reativas de Oxigênio/metabolismoRESUMO
Industrial wine yeasts owe their adaptability in constantly changing environments to a long evolutionary history that combines naturally occurring evolutionary events with human-enforced domestication. Among the many stressors associated with winemaking processes that have potentially detrimental impacts on yeast viability, growth, and fermentation performance are hyperosmolarity, high glucose concentrations at the beginning of fermentation, followed by the depletion of nutrients at the end of this process. Therefore, in this study, we subjected three widely used industrial wine yeasts to adaptive laboratory evolution under potassium chloride (KCl)-induced osmotic stress. At the end of the evolutionary experiment, we evaluated the tolerance to high osmotic stress of the evolved strains. All of the analyzed strains improved their fitness under high osmotic stress without worsening their economic characteristics, such as growth rate and viability. The evolved derivatives of two strains also gained the ability to accumulate glycogen, a readily mobilized storage form of glucose conferring enhanced viability and vitality of cells during prolonged nutrient deprivation. Moreover, laboratory-scale fermentation in grape juice showed that some of the KCl-evolved strains significantly enhanced glycerol synthesis and production of resveratrol-enriched wines, which in turn greatly improved the wine sensory profile. Altogether, these findings showed that long-term adaptations to osmotic stress can be an attractive approach to develop industrial yeasts.
Assuntos
Adaptação Fisiológica/genética , Pressão Osmótica/fisiologia , Saccharomyces cerevisiae/fisiologia , Vitis/fisiologia , Etanol/metabolismo , Fermentação , Glucose/metabolismo , Glicogênio/metabolismo , Humanos , Vinho/microbiologia , Fermento Seco/genéticaRESUMO
Matcha green tea (Camellia sinensis), which originates from Japan, is commonly considered as particularly beneficial to health. A large content of polyphenols, amino acids (mainly tannins) and caffeine potentially increase the antioxidant properties of the drink. The aim of the study was to determine the antioxidant potential and the content of substances with an antioxidant effect-vitamin C, total polyphenol content including flavonoids-in infusions made from Traditional Matcha (from the first and second harvests) and Daily Matcha (from the second and third harvests) at different temperatures. The infusions were made by pouring 100 mL of distilled water once at various temperatures (25 °C, 70 °C, 80 °C and 90 °C) over 1.75 g of the plant material. Matcha tea is characterized by a high level of antioxidant substances (flavonoids 1968.8 mg/L; polyphenols 1765.1 mg/L; vitamin C 44.8 mg/L) as well as antioxidant potential (41.2% DPPH (10× dilution); 6129.5 µM Fe(II)/dm3 FRAP). The concentration of these compounds depends on the time at which the plant material was harvested as well as on the temperature of water used to prepare the infusions. For most parameters, the highest values were observed in infusions prepared at 90 °C and from the daily Matcha.
RESUMO
Distillery yeasts are poorly characterized physiological group among the Saccharomyces sensu stricto complex. As industrial yeasts are under constant environmental stress during fermentation processes and the nucleolus is a stress sensor, in the present study, nucleolus-related parameters were evaluated in 22 commercially available distillery yeast strains. Distillery yeasts were found to be a heterogeneous group with a variable content and length of rDNA and degree of nucleolus fragmentation. The levels of rDNA were negatively correlated with Nop1 (r = -0.59, p = 0.0038). Moreover, the protein levels of Sir transcriptional silencing complex and longevity regulators, namely Sir1, Sir2, Sir3 and Fob1, were studied and negative correlations between Sir2 and Nop1 (r = -0.45, p = 0.0332), and between Sir2 and Fob1 (r = -0.49, p = 0.0211) were revealed. In general, S. paradoxus group of distillery yeasts with higher rDNA pools and Sir2 level than S. bayanus group was found to be more tolerant to fermentation-associated stress stimuli, namely mild cold/heat stresses and KCl treatment. We postulate that rDNA state may be considered as a novel factor that may modulate a biotechnological process.
Assuntos
Bebidas Alcoólicas/microbiologia , DNA Ribossômico/genética , Proteínas Nucleares/genética , Ribonucleoproteínas Nucleolares Pequenas/genética , Proteínas de Saccharomyces cerevisiae/genética , Saccharomyces cerevisiae/genética , Proteínas Reguladoras de Informação Silenciosa de Saccharomyces cerevisiae/genética , Sirtuína 2/genética , Proteínas de Ligação a DNA/genética , Região Organizadora do Nucléolo/genéticaRESUMO
The yeast strains of the Saccharomyces sensu stricto complex involved in beer production are a heterogeneous group whose genetic and genomic features are not adequately determined. Thus, the aim of the present study was to provide a genetic characterization of selected group of commercially available brewing yeasts both ale top-fermenting and lager bottom-fermenting strains. Molecular karyotyping revealed that the diversity of chromosome patterns and four strains with the most accented genetic variabilities were selected and subjected to genome-wide array-based comparative genomic hybridization (array-CGH) analysis. The differences in the gene copy number were found in five functional gene categories: (1) maltose metabolism and transport, (2) response to toxin, (3) siderophore transport, (4) cellular aldehyde metabolic process, and (5) L-iditol 2-dehydrogenase activity (p < 0.05). In the Saflager W-34/70 strain (Fermentis) with the most affected array-CGH profile, loss of aryl-alcohol dehydrogenase (AAD) gene dosage correlated with an imbalanced redox state, oxidative DNA damage and breaks, lower levels of nucleolar proteins Nop1 and Fob1, and diminished tolerance to fermentation-associated stress stimuli compared to other strains. We suggest that compromised stress response may not only promote oxidant-based changes in the nucleolus state that may affect fermentation performance but also provide novel directions for future strain improvement.
Assuntos
Variações do Número de Cópias de DNA , Saccharomyces cerevisiae/genética , Estresse Fisiológico , Nucléolo Celular/fisiologia , Cromossomos Fúngicos/genética , Dano ao DNA , Fermentação , Ontologia Genética , Genes Fúngicos , Instabilidade Genômica , Viabilidade Microbiana , Oxirredução , Estresse Oxidativo , Ploidias , Saccharomyces cerevisiae/crescimento & desenvolvimentoRESUMO
Industrial yeast strains of economic importance used in winemaking and beer production are genomically diverse and subjected to harsh environmental conditions during fermentation. In the present study, we investigated wine yeast adaptation to chronic mild alcohol stress when cells were cultured for 100 generations in the presence of non-cytotoxic ethanol concentration. Ethanol-induced reactive oxygen species (ROS) and superoxide signals promoted growth rate during passages that was accompanied by increased expression of sirtuin proteins, Sir1, Sir2 and Sir3, and DNA-binding transcription regulator Rap1. Genome-wide array-CGH analysis revealed that yeast genome was shaped during passages. The gains of chromosomes I, III and VI and significant changes in the gene copy number in nine functional gene categories involved in metabolic processes and stress responses were observed. Ethanol-mediated gains of YRF1 and CUP1 genes were the most accented. Ethanol also induced nucleolus fragmentation that confirms that nucleolus is a stress sensor in yeasts. Taken together, we postulate that wine yeasts of different origin may adapt to mild alcohol stress by shifts in intracellular redox state promoting growth capacity, upregulation of key regulators of longevity, namely sirtuins and changes in the dosage of genes involved in the telomere maintenance and ion detoxification.
Assuntos
Adaptação Biológica/efeitos dos fármacos , Cromossomos Fúngicos/genética , Etanol/farmacologia , Fermentação/fisiologia , Espécies Reativas de Oxigênio/metabolismo , Saccharomyces cerevisiae/fisiologia , Cerveja , Nucléolo Celular/efeitos dos fármacos , Cromossomos Fúngicos/efeitos dos fármacos , Hibridização Genômica Comparativa , Indústria Alimentícia , Dosagem de Genes , Oxirredução , Proteínas de Saccharomyces cerevisiae/metabolismo , Complexo Shelterina , Transdução de Sinais/efeitos dos fármacos , Proteínas Reguladoras de Informação Silenciosa de Saccharomyces cerevisiae/metabolismo , Sirtuína 2/metabolismo , Sirtuínas , Homeostase do Telômero/efeitos dos fármacos , Homeostase do Telômero/genética , Proteínas de Ligação a Telômeros/metabolismo , Fatores de Transcrição/metabolismo , VinhoRESUMO
The aim of the present study was to investigate the effects of crossbreeding on collagen content and solubility, shear force (WBSF) and the eating quality of Infraspinatus (INF) and Semimembranosus (SM) muscles of young bulls and the relationships between collagen content and solubility, shear force and the eating quality of beef. The experimental material comprised muscles of crossbred young bulls (about 600 days old) of Polish Holstein-Friesian (PHF) × Limousine (LM) (n = 10), PHF × Charolaise (CH) (n = 9), PHF × Hereford (HER) (n = 9) breeds. The crossbreeding influenced WBSF, aroma and taste, total, water-soluble, acid-soluble, total soluble and insoluble collagen content, as well as the acid-soluble, total soluble and insoluble collagen proportions. WBSF was significantly negatively correlated with sensorial tenderness and water-soluble collagen content. The eating quality of beef obtained from different crossbreds was similar; however, the meat from PHF × LM and PHF × HER bulls had lower WBSF values than PHF × CH bulls. © 2016 Japanese Society of Animal Science.
Assuntos
Bovinos/genética , Colágeno/análise , Qualidade dos Alimentos , Hibridização Genética , Carne , Músculo Esquelético , Característica Quantitativa Herdável , Solubilidade , Animais , Humanos , Masculino , Carne/análise , Resistência ao Cisalhamento , Paladar , ÁguaRESUMO
The nucleolus is considered to be a stress sensor and rDNA-based regulation of cellular senescence and longevity has been proposed. However, the role of rDNA in the maintenance of genome integrity has not been investigated in detail. Using genomically diverse industrial yeasts as a model and array-based comparative genomic hybridization (aCGH), we show that chromosome level may be balanced during passages and as a response to alcohol stress that may be associated with changes in rDNA pools. Generation- and ethanol-mediated changes in genes responsible for protein and DNA/RNA metabolism were revealed using next-generation sequencing. Links between redox homeostasis, DNA stability, and telomere and nucleolus states were also established. These results suggest that yeast genome is dynamic and chromosome homeostasis may be controlled by rDNA.
Assuntos
Nucléolo Celular/genética , Cromossomos Fúngicos/genética , DNA Fúngico/genética , DNA Ribossômico/genética , Genoma Fúngico , Saccharomyces cerevisiae/genética , Nucléolo Celular/efeitos dos fármacos , Nucléolo Celular/metabolismo , Cromossomos Fúngicos/metabolismo , Hibridização Genômica Comparativa , Dano ao DNA , DNA Fúngico/metabolismo , DNA Ribossômico/metabolismo , Etanol/toxicidade , Regulação Fúngica da Expressão Gênica , Instabilidade Genômica , Homeostase , Oxirredução , Estresse Oxidativo , Saccharomyces cerevisiae/efeitos dos fármacos , Saccharomyces cerevisiae/crescimento & desenvolvimento , Saccharomyces cerevisiae/metabolismo , Seleção Genética , Telômero/genética , Telômero/metabolismoRESUMO
Industrial yeasts, economically important microorganisms, are widely used in diverse biotechnological processes including brewing, winemaking and distilling. In contrast to a well-established genome of brewer's and wine yeast strains, the comprehensive evaluation of genomic features of distillery strains is lacking. In the present study, twenty two distillery yeast strains were subjected to electrophoretic karyotyping and array-based comparative genomic hybridization (array-CGH). The strains analyzed were assigned to the Saccharomyces sensu stricto complex and grouped into four species categories: S. bayanus, S. paradoxus, S. cerevisiae and S. kudriavzevii. The genomic diversity was mainly revealed within subtelomeric regions and the losses and/or gains of fragments of chromosomes I, III, VI and IX were the most frequently observed. Statistically significant differences in the gene copy number were documented in six functional gene categories: 1) telomere maintenance via recombination, DNA helicase activity or DNA binding, 2) maltose metabolism process, glucose transmembrane transporter activity; 3) asparagine catabolism, cellular response to nitrogen starvation, localized in cell wall-bounded periplasmic space, 4) siderophore transport, 5) response to copper ion, cadmium ion binding and 6) L-iditol 2- dehydrogenase activity. The losses of YRF1 genes (Y' element ATP-dependent helicase) were accompanied by decreased level of Y' sequences and an increase in DNA double and single strand breaks, and oxidative DNA damage in the S. paradoxus group compared to the S. bayanus group. We postulate that naturally occurring diversity in the YRF1 gene copy number may promote genetic stability in the S. bayanus group of distillery yeast strains.
Assuntos
Variações do Número de Cópias de DNA/genética , DNA Helicases/genética , Dosagem de Genes/genética , Proteínas de Saccharomyces cerevisiae/genética , Saccharomyces/genética , Asparagina/metabolismo , Hibridização Genômica Comparativa , Quebras de DNA de Cadeia Dupla , Quebras de DNA de Cadeia Simples , Variação Genética/genética , Cariotipagem , L-Iditol 2-Desidrogenase/metabolismo , Oxirredução , Saccharomyces/classificação , Vinho/microbiologiaRESUMO
Changes in the water holding capacity (WHC) and collagen profile in bovine m. infraspinatus during ageing in a vacuum at 3 °C up to 20 days were studied. Moisture, expressible water and total collagen content were determined in raw samples and muscles were then subjected to thermal treatment in steam. Moisture, collagen fractions (total, water-soluble, insoluble, acetic acid-soluble, total soluble) and cooking loss were determined in the cooked meat. Ageing time did not significantly influence WHC (expressible water content as a % of moisture), moisture content in raw and cooked meat or cooking loss. Between the 5th and 10th day of ageing a significant increase was noted in acetic acid-soluble collagen and total soluble collagen content along with a decrease in insoluble collagen content (p b 0.05). A significant negative relationship was found between the expressible water content and the water-soluble collagen. It was concluded that connective tissue ageing of bovine m. infraspinatus might be finished after 10 days.
Assuntos
Colágeno/análise , Tecido Conjuntivo/química , Carne/análise , Mudanças Depois da Morte , Água/análise , Ácido Acético , Animais , Bovinos , Culinária , Manipulação de Alimentos , Temperatura Alta , Humanos , Músculo Esquelético , Solubilidade , Vapor , VácuoRESUMO
Plant cell suspension cultures represent good model systems applicable for both basic research and biotechnological purposes. Nevertheless, it is widely known that a prolonged in vitro cultivation of plant cells is associated with genetic and epigenetic instabilities, which may limit the usefulness of plant lines. In this study, the age-dependent epigenetic and physiological changes in an asynchronous Arabidopsis T87 cell culture were examined. A prolonged cultivation period was found to be correlated with a decrease in the proliferation rate and a simultaneous increase in the expression of senescence-associated genes, indicating that the aging process started at the late growth phase of the culture. In addition, increases in the heterochromatin-specific epigenetic markers, i.e., global DNA methylation, H3K9 dimethylation, and H3K27 trimethylation, were observed, suggesting the onset of chromatin condensation, a hallmark of the early stages of plant senescence. Although the number of live cells decreased with an increase in the age of the culture, the remaining viable cells retained a high potential to efficiently perform photosynthesis and did not exhibit any symptoms of photosystem II damage.
Assuntos
Arabidopsis/citologia , Senescência Celular/genética , Senescência Celular/fisiologia , Epigênese Genética , Arabidopsis/genética , Arabidopsis/fisiologia , Técnicas de Cultura de Células , Linhagem Celular , Proliferação de Células , Sobrevivência Celular , Clorofila/análise , Clorofila A , Metilação de DNA , Fluorescência , Expressão Gênica , Heterocromatina/química , Heterocromatina/metabolismo , Suspensões , Fatores de TempoRESUMO
A genomic in situ hybridization (GISH)-based method for dermatophyte identification has been developed. Using specific GISH probes, discrimination between Trichophyton interdigitale, Trichophyton rubrum and Microsporum canis has been conducted. Moreover, GISH has been found particularly helpful when proper dermatophyte identification was difficult due to ambiguous PCR-RFLP patterns.
Assuntos
Arthrodermataceae/genética , Arthrodermataceae/isolamento & purificação , Dermatomicoses/diagnóstico , Hibridização In Situ/métodos , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Animais , Arthrodermataceae/classificação , Criança , Pré-Escolar , Dermatomicoses/microbiologia , Feminino , Genoma Fúngico , Humanos , Masculino , Microsporum/classificação , Microsporum/genética , Microsporum/isolamento & purificação , Pessoa de Meia-Idade , Trichophyton/classificação , Trichophyton/genética , Trichophyton/isolamento & purificação , Adulto JovemRESUMO
The aim of the study was to evaluate the influence of cooking conditions (dry air and steam) and final internal temperature (75, 85, 95°C) on the physico-chemical properties of beef infraspinatus (INF) and semimembranosus (SEM) muscles as well as their tenderness and juiciness. Cooking method and temperature influenced moisture, total collagen content in cooked meat and cooking loss, whereas muscle type affected fat, total collagen content and cooking loss. Warner-Bratzler shear force values were affected by cooking method, which also influenced juiciness of roasts. Temperature affected tenderness and juiciness, whereas muscle type influenced juiciness. The most desirable tenderness had INF heated in steam and dry air to 95°C. Processing SEM in dry air to 85 and 95°C lowered the juiciness of the roasts. There were significant correlations between physico-chemical, sensorial and image attributes, however high accuracy of prediction (r(2)>0.8) was achieved only for SEM muscle.
Assuntos
Culinária/métodos , Carne/análise , Músculo Esquelético/química , Temperatura , Animais , Água Corporal , Bovinos , Colágeno/análise , Gorduras/análise , Tecnologia de Alimentos , Temperatura Alta , Humanos , Carne/normas , Estresse MecânicoRESUMO
Animal cells react to mitogenic or stress stimuli by rapid up-regulation of immediate-early (IE) genes and a parallel increase in characteristic modifications of core histones: chromatin changes, collectively termed the nucleosomal response. With regard to plants little is known about the accompanying changes at the chromatin level. We have used tobacco BY-2 and Arabidopsis T87 cell lines to study the nucleosomal response of plant cells to high salinity, cold and exogenous abscisic acid (ABA). When in quiescent stage, both tobacco and Arabidopsis cells show the typical nucleosomal response to high salinity and cold stress, manifested by rapid transient up-regulation of histone H3 Ser-10 phosphorylation, immediately followed by transient up-regulation of H3 phosphoacetylation and histone H4 acetylation. For each of the studied stresses the observed nucleosomal response was strictly correlated with the induction of stress-type specific genes. The dynamics of histone modifications in BY-2 cells in response to exogenous ABA exhibited a more complex pattern than that evoked by the two abiotic stresses, probably due to superposition of the primary and secondary effects of ABA. A rapid increase in H3 Ser-10 phosphorylation was also observed in whole leaves subjected to high salinity; however, the rate of change in this modification was much slower than in cultured cells. Together, these results indicate that the quiescent BY-2 and T87 cell lines show a typical nucleosomal response to abiotic stresses and ABA treatment and may represent suitable models for the study of chromatin-mediated mechanisms of stress tolerance in plants.
Assuntos
Ácido Abscísico/farmacologia , Arabidopsis/genética , Genes de Plantas , Histonas/metabolismo , Nicotiana/genética , Regulação para Cima/efeitos dos fármacos , Acetilação/efeitos dos fármacos , Arabidopsis/citologia , Arabidopsis/metabolismo , Western Blotting , Células Cultivadas , Eletroforese em Gel de Poliacrilamida , Regulação da Expressão Gênica de Plantas/efeitos dos fármacos , Nucleossomos/metabolismo , Fosforilação/efeitos dos fármacos , Reguladores de Crescimento de Plantas/farmacologia , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Serina/metabolismo , Cloreto de Sódio/farmacologia , Nicotiana/citologia , Nicotiana/metabolismo , Regulação para Cima/genéticaRESUMO
SWITCH/SUCROSE NONFERMENTING (SWI/SNF) chromatin-remodeling complexes mediate ATP-dependent alterations of DNA-histone contacts. The minimal functional core of conserved SWI/SNF complexes consists of a SWI2/SNF2 ATPase, SNF5, SWP73, and a pair of SWI3 subunits. Because of early duplication of the SWI3 gene family in plants, Arabidopsis thaliana encodes four SWI3-like proteins that show remarkable functional diversification. Whereas ATSWI3A and ATSWI3B form homodimers and heterodimers and interact with BSH/SNF5, ATSWI3C, and the flowering regulator FCA, ATSWI3D can only bind ATSWI3B in yeast two-hybrid assays. Mutations of ATSWI3A and ATSWI3B arrest embryo development at the globular stage. By a possible imprinting effect, the atswi3b mutations result in death for approximately half of both macrospores and microspores. Mutations in ATSWI3C cause semidwarf stature, inhibition of root elongation, leaf curling, aberrant stamen development, and reduced fertility. Plants carrying atswi3d mutations display severe dwarfism, alterations in the number and development of flower organs, and complete male and female sterility. These data indicate that, by possible contribution to the combinatorial assembly of different SWI/SNF complexes, the ATSWI3 proteins perform nonredundant regulatory functions that affect embryogenesis and both the vegetative and reproductive phases of plant development.