RESUMO
Artemisia argyi is an important medicinal and economic plant in China, with the effects of warming channels, dispersing cold, and relieving pain, inflammation, and allergy. The essential oil of this plant is rich in volatile terpenoids and widely used in moxi-bustion and healthcare products, with huge market potential. The bZIP transcription factors compose a large family in plants and are involved in the regulation of plant growth and development, stress response, and biosynthesis of secondary metabolites such as terpenoids. However, little is known about the bZIPs and their roles in A. argyi. In this study, the bZIP transcription factors in the genome of A. argyi were systematically identified, and their physicochemical properties, phylogenetic relationship, conserved motifs, and promoter-binding elements were analyzed. Candidate AarbZIP genes involved in terpenoid biosynthesis were screened out. The results showed that a total of 156 AarbZIP transcription factors were identified at the genomic level, with the lengths of 99-618 aa, the molecular weights of 11.7-67.8 kDa, and the theoretical isoelectric points of 4.56-10.16. According to the classification of bZIPs in Arabidopsis thaliana, the 156 AarbZIPs were classified into 12 subfamilies, and the members in the same subfamily had similar conserved motifs. The cis-acting elements of promoters showed that AarbZIP genes were possibly involved in light and hormonal pathways. Five AarbZIP genes that may be involved in the regulation of terpenoid biosynthesis were screened out by homologous alignment and phylogenetic analysis. The qRT-PCR results showed that the expression levels of the five AarbZIP genes varied significantly in different tissues of A. argyi. Specifically, AarbZIP29 and AarbZIP55 were highly expressed in the leaves and AarbZIP81, AarbZIP130, and AarbZIP150 in the flower buds. This study lays a foundation for the functional study of bZIP genes and their regulatory roles in the terpenoid biosynthesis in A. argyi.
Assuntos
Artemisia , Perfilação da Expressão Gênica , Filogenia , Artemisia/genética , Fatores de Transcrição de Zíper de Leucina Básica/genética , Fatores de Transcrição de Zíper de Leucina Básica/metabolismo , Terpenos , Regulação da Expressão Gênica de PlantasRESUMO
MAIN CONCLUSION: This study provides an overview of the structure, classification, regulatory mechanisms, and biological functions of the basic (region) leucine zipper transcription factors and their molecular mechanisms in flavonoid, terpenoid, alkaloid, phenolic acid, and lignin biosynthesis. Basic (region) leucine zippers (bZIPs) are evolutionarily conserved transcription factors (TFs) in eukaryotic organisms. The bZIP TFs are widely distributed in plants and play important roles in plant growth and development, photomorphogenesis, signal transduction, resistance to pathogenic microbes, biotic and abiotic stress, and secondary metabolism. Moreover, the expression of bZIP TFs not only promotes or inhibits the accumulation of secondary metabolites in medicinal plants, but also affects the stress response of plants to the external adverse environment. This paper describes the structure, classification, biological function, and regulatory mechanisms of bZIP TFs. In addition, the molecular mechanism of bZIP TFs regulating the biosynthesis of flavonoids, terpenoids, alkaloids, phenolic acids, and lignin are also elaborated. This review provides a summary for in-depth study of the molecular mechanism of bZIP TFs regulating the synthesis pathway of secondary metabolites and plant molecular breeding, which is of significance for the generation of beneficial secondary metabolites and the improvement of plant varieties.
Assuntos
Fatores de Transcrição de Zíper de Leucina Básica , Lignina , Fatores de Transcrição de Zíper de Leucina Básica/genética , Fatores de Transcrição de Zíper de Leucina Básica/metabolismo , Metabolismo Secundário/genética , Lignina/metabolismo , Plantas/genética , Plantas/metabolismo , Fatores de Transcrição/genética , Fatores de Transcrição/metabolismo , Estresse Fisiológico , Regulação da Expressão Gênica de Plantas , Proteínas de Plantas/metabolismo , FilogeniaRESUMO
BACKGROUND: The bZIP gene family has important roles in various biological processes, including development and stress responses. However, little information about this gene family is available for Wheel Wingnut (Cyclocarya paliurus). RESULTS: In this study, we identified 58 bZIP genes in the C. paliurus genome and analyzed phylogenetic relationships, chromosomal locations, gene structure, collinearity, and gene expression profiles. The 58 bZIP genes could be divided into 11 groups and were unevenly distributed among 16 C. paliurus chromosomes. An analysis of cis-regulatory elements indicated that bZIP promoters were associated with phytohormones and stress responses. The expression patterns of bZIP genes in leaves differed among developmental stages. In addition, several bZIP members were differentially expressed under drought stress. These expression patterns were verified by RT-qPCR. CONCLUSIONS: Our results provide insights into the evolutionary history of the bZIP gene family in C. paliurus and the function of these genes during leaf development and in the response to drought stress. In addition to basic genomic information, our results provide a theoretical basis for further studies aimed at improving growth and stress resistance in C. paliurus, an important medicinal plant.
Assuntos
Secas , Regulação da Expressão Gênica de Plantas , Filogenia , Fatores de Transcrição de Zíper de Leucina Básica/genética , Fatores de Transcrição de Zíper de Leucina Básica/metabolismo , Perfilação da Expressão GênicaRESUMO
As one of the largest transcription factor families in plants, bZIP transcription factors play important regulatory roles in different biological processes, especially in the process of stress response. Salt stress inhibits the growth and yield of sugar beet. However, bZIP-related studies in sugar beet (Beta vulgaris L.) have not been reported. This study aimed to identify the bZIP transcription factors in sugar beet and analyze their biological functions and response patterns to salt stress. Using bioinformatics, 48 BvbZIP genes were identified in the genome of sugar beet, encoding 77 proteins with large structural differences. Collinearity analysis showed that three pairs of BvbZIP genes were fragment replication genes. The BvbZIP genes were grouped according to the phylogenetic tree topology and conserved structures, and the results are consistent with those reported in Arabidopsis. Under salt stress, the expression levels of most BvbZIP genes were decreased, and only eight genes were up-regulated. GO analysis showed that the BvbZIP genes were mainly negatively regulated in stress response. Protein interaction prediction showed that the BvbZIP genes were mainly involved in light signaling and ABA signal transduction, and also played a certain role in stress responses. In this study, the structures and biological functions of the BvbZIP genes were analyzed to provide foundational data for further mechanistic studies and for facilitating the efforts toward the molecular breeding of stress-resilient sugar beet.
Assuntos
Arabidopsis , Beta vulgaris , Arabidopsis/genética , Arabidopsis/metabolismo , Fatores de Transcrição de Zíper de Leucina Básica/genética , Fatores de Transcrição de Zíper de Leucina Básica/metabolismo , Regulação da Expressão Gênica de Plantas , Filogenia , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Estresse Salino/genética , Estresse Fisiológico/genética , Açúcares/metabolismo , Fatores de Transcrição/metabolismoRESUMO
Exposure to lead (Pb), a known toxin causing developmental neurotoxicity, can impair neurogenesis and oxidative phosphorylation (OXPHOS), but the mechanism is not clarified. In the current study, we aim to explore the effects of Pb on the differentiation of SH-SY5Y cells and investigate the role of heme and heme-binding protein BACH1 during differentiation. We found that Pb exposure caused a shift from OXPHOS to glycolysis, resulting in neurogenesis impairment by decreasing neurite growth and downregulation of PSD95 and Synapsin-1 in differentiated SH-SY5Y cells. Heme reduction mediated this mitochondria metabolism repression caused by Pb depending on BACH1 activation. Hemin supplement alleviated Pb-induced OXPHOS damage and adenosine triphosphate (ATP) reduction in differentiated SH-SY5Y cells, and further protected for Pb-induced damage of synapse. Heme binding factor BACH1 was negatively regulated by heme content and BACH1 knockout rescued the Pb-induced transcription and expression decline of genes related to OXPHOS and abrogated Pb-induced growth inhibition of axon promotion and synapse formation. Collectively, the present study demonstrates that heme deficiency mediates OXPHOS damage caused by Pb through BACH1 activation, resulting in neurogenesis impairment.
Assuntos
Hemina , Neuroblastoma , Humanos , Hemina/metabolismo , Hemina/farmacologia , Chumbo/toxicidade , Chumbo/metabolismo , Proteínas Ligantes de Grupo Heme , Sinapsinas/metabolismo , Sinapsinas/farmacologia , Neuroblastoma/metabolismo , Mitocôndrias , Heme/metabolismo , Trifosfato de Adenosina/metabolismo , Linhagem Celular Tumoral , Fatores de Transcrição de Zíper de Leucina Básica/genética , Fatores de Transcrição de Zíper de Leucina Básica/metabolismo , Fatores de Transcrição de Zíper de Leucina Básica/farmacologiaRESUMO
Basic leucine zipper (bZIP) transcription factors play an irreplaceable position in the regulation of plant secondary metabolism, growth and development, and resistance to abiotic stress. Panax notoginseng is a traditional medicinal plant in China, but the systematic identification and the resistance of Panax notoginseng bZIP (PnbZIP) family under nitrogen stress have not been reported before, considering the excessive application of N fertilizers. In this study, we conducted a genome-wide identification of the PnbZIP family and analyzed its phylogeny, tissue selectivity, and abiotic resistence. 74 PnbZIPs were distributed on 12 chromosomes and 8 were not successfully located. Through phylogenetic analysis of Arabidopsis and Panax notoginseng, we divided them into 14 subgroups. In the same subgroup, bZIPs had similiar intron/exon structure and conserved motifs. In the analysis of chromosome structure, two PnbZIP genes were duplicated in tandem on chromosome 3. Intraspecific collinearity analysis showed that 28 PnbZIPs participated in segmental replication. Each PnbZIP promoter contained at least one stress response element or stress-related hormone response element. RNA-seq and qRT-PCR methods were used to analyze the expression patterns of the PnbZIP gene in different tissues (roots, flowers, and leaves) and under different nitrogen stresses. The results showed that the PnbZIP gene had the highest expression level in flowers and reflected tissue-specific expressions. Meanwhile, under the stress of ammonium nitrogen fertilizer and nitrate nitrogen fertilizer, PnbZIPs in roots were differently expressed. 10 PnbZIP stress-responsive genes were screened for significant expression, among which PnbZIP46 was significantly up-regulated, which could be a candidate gene for resistance to Nitrogen stress. This study laid the foundation for functional identification of PnbZIPs and improved the cultivation of Panax notoginseng.
Assuntos
Regulação da Expressão Gênica de Plantas , Panax notoginseng , Algoritmos , Fatores de Transcrição de Zíper de Leucina Básica/genética , Fatores de Transcrição de Zíper de Leucina Básica/metabolismo , Fertilizantes , Perfilação da Expressão Gênica , Nitrogênio/metabolismo , Nitrogênio/farmacologia , Panax notoginseng/genética , Panax notoginseng/metabolismo , Filogenia , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Estresse Fisiológico/genéticaRESUMO
HAC1 mRNA remains translationally repressed in the cytoplasm of the budding yeast Saccharomyces cerevisiae. Under conditions of cellular stress, a dual kinase RNase IRE1 (Inositol Requiring Enzyme-1) cleaves out an intervening sequence from the HAC1 mRNA. Cleaved mRNAs are then ligated by tRNA ligase, thus generating a spliced mRNA that translates an active transcription factor. This unconventional splicing of HAC1 mRNA in the cytoplasm is a molecular marker for various cellular stresses including oxidative stress and endoplasmic reticulum (ER) stress. This article describes a PCR-based protocol to detect the HAC1 mRNA splicing.
Assuntos
Proteínas de Saccharomyces cerevisiae , Saccharomyces cerevisiae , Fatores de Transcrição de Zíper de Leucina Básica/genética , Fatores de Transcrição de Zíper de Leucina Básica/metabolismo , Regulação Fúngica da Expressão Gênica , Splicing de RNA , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Proteínas Repressoras/genética , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Saccharomyces cerevisiae/genética , Saccharomyces cerevisiae/metabolismo , Proteínas de Saccharomyces cerevisiae/genética , Proteínas de Saccharomyces cerevisiae/metabolismoRESUMO
Type 2 diabetes (T2D) is associated with defective insulin secretion and reduced ß cell mass. Available treatments provide a temporary reprieve, but secondary failure rates are high, making insulin supplementation necessary. Reversibility of ß cell failure is a key translational question. Here, we reverse engineered and interrogated pancreatic islet-specific regulatory networks to discover T2D-specific subpopulations characterized by metabolic inflexibility and endocrine progenitor/stem cell features. Single-cell gain- and loss-of-function and glucose-induced Ca2+ flux analyses of top candidate master regulatory (MR) proteins in islet cells validated transcription factor BACH2 and associated epigenetic effectors as key drivers of T2D cell states. BACH2 knockout in T2D islets reversed cellular features of the disease, restoring a nondiabetic phenotype. BACH2-immunoreactive islet cells increased approximately 4-fold in diabetic patients, confirming the algorithmic prediction of clinically relevant subpopulations. Treatment with a BACH inhibitor lowered glycemia and increased plasma insulin levels in diabetic mice, and restored insulin secretion in diabetic mice and human islets. The findings suggest that T2D-specific populations of failing ß cells can be reversed and indicate pathways for pharmacological intervention, including via BACH2 inhibition.
Assuntos
Fatores de Transcrição de Zíper de Leucina Básica/antagonistas & inibidores , Fatores de Transcrição de Zíper de Leucina Básica/metabolismo , Sinalização do Cálcio , Diabetes Mellitus Tipo 2/metabolismo , Epigênese Genética , Células Secretoras de Insulina/metabolismo , Fatores de Transcrição de Zíper de Leucina Básica/genética , Diabetes Mellitus Tipo 2/tratamento farmacológico , Diabetes Mellitus Tipo 2/genética , Células HEK293 , HumanosRESUMO
Parkinson's disease (PD) is a progressive neurodegenerative movement disorder characterized by the loss of nigrostriatal dopaminergic neurons. Mounting evidence suggests that Nrf2 is a promising target for neuroprotective interventions in PD. However, electrophilic chemical properties of the canonical Nrf2-based drugs cause irreversible alkylation of cysteine residues on cellular proteins resulting in side effects. Bach1 is a known transcriptional repressor of the Nrf2 pathway. We report that Bach1 levels are up-regulated in PD postmortem brains and preclinical models. Bach1 knockout (KO) mice were protected against 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP)-induced dopaminergic neurotoxicity and associated oxidative damage and neuroinflammation. Functional genomic analysis demonstrated that the neuroprotective effects in Bach1 KO mice was due to up-regulation of Bach1-targeted pathways that are associated with both Nrf2-dependent antioxidant response element (ARE) and Nrf2-independent non-ARE genes. Using a proprietary translational technology platform, a drug library screen identified a substituted benzimidazole as a Bach1 inhibitor that was validated as a nonelectrophile. Oral administration of the Bach1 inhibitor attenuated MPTP neurotoxicity in pre- and posttreatment paradigms. Bach1 inhibitor-induced neuroprotection was associated with the up-regulation of Bach1-targeted pathways in concurrence with the results from Bach1 KO mice. Our results suggest that genetic deletion as well as pharmacologic inhibition of Bach1 by a nonelectrophilic inhibitor is a promising therapeutic approach for PD.
Assuntos
Fatores de Transcrição de Zíper de Leucina Básica/metabolismo , Neuroproteção , Doença de Parkinson/terapia , 1-Metil-4-Fenil-1,2,3,6-Tetra-Hidropiridina , Idoso , Idoso de 80 Anos ou mais , Animais , Elementos de Resposta Antioxidante , Fatores de Transcrição de Zíper de Leucina Básica/antagonistas & inibidores , Fatores de Transcrição de Zíper de Leucina Básica/genética , Estudos de Casos e Controles , Modelos Animais de Doenças , Avaliação Pré-Clínica de Medicamentos , Feminino , Humanos , Masculino , Camundongos , Camundongos Knockout , Doença de Parkinson/metabolismo , RatosRESUMO
The central nervous system regulates activity of peripheral organs through interoception. In our previous study, we have demonstrated that PGE2/EP4 skeleton interception regulate bone homeostasis. Here, we show that ascending skeleton interoceptive signaling downregulates expression of hypothalamic neuropeptide Y (NPY) and induce lipolysis of adipose tissue for osteoblastic bone formation. Specifically, the ascending skeleton interoceptive signaling induces expression of small heterodimer partner-interacting leucine zipper protein (SMILE) in the hypothalamus. SMILE binds to pCREB as a transcriptional heterodimer on Npy promoters to inhibit NPY expression. Knockout of EP4 in sensory nerve increases expression of NPY causing bone catabolism and fat anabolism. Importantly, inhibition of NPY Y1 receptor (Y1R) accelerated oxidation of free fatty acids in osteoblasts and rescued bone loss in AvilCre:Ptger4fl/fl mice. Thus, downregulation of hypothalamic NPY expression lipolyzes free fatty acids for anabolic bone formation through a neuroendocrine descending interoceptive regulation.
Assuntos
Tecido Adiposo/metabolismo , Osso e Ossos/metabolismo , Hipotálamo/fisiologia , Interocepção/fisiologia , Neuropeptídeo Y/metabolismo , Esqueleto/fisiologia , Animais , Fatores de Transcrição de Zíper de Leucina Básica/genética , Fatores de Transcrição de Zíper de Leucina Básica/metabolismo , Regulação para Baixo , Regulação da Expressão Gênica , Metabolismo dos Lipídeos , Camundongos , Camundongos Knockout , Neuropeptídeo Y/genética , Osteoblastos/fisiologia , Transdução de SinaisRESUMO
Radiotherapy, the most common therapy for the treatment of solid tumors, exerts its effects by inducing DNA damage. To fully understand the extent and nature of this damage, DNA models that mimic the in vivo situation should be utilized. In a cellular context, genomic DNA constantly interacts with proteins and these interactions could influence both the primary radical processes (triggered by ionizing radiation) and secondary reactions, ultimately leading to DNA damage. However, this is seldom addressed in the literature. In this work, we propose a general approach to tackle these shortcomings. We synthesized a protein-DNA complex that more closely represents DNA in the physiological environment than oligonucleotides solution itself, while being sufficiently simple to permit further chemical analyses. Using click chemistry, we obtained an oligonucleotide-peptide conjugate, which, if annealed with the complementary oligonucleotide strand, forms a complex that mimics the specific interactions between the GCN4 protein and DNA. The covalent bond connecting the oligonucleotide and peptide constitutes a part of substituted triazole, which forms due to the click reaction between the short peptide corresponding to the specific amino acid sequence of GCN4 protein (yeast transcription factor) and a DNA fragment that is recognized by the protein. DNAse footprinting demonstrated that the part of the DNA fragment that specifically interacts with the peptide in the complex is protected from DNAse activity. Moreover, the thermodynamic characteristics obtained using differential scanning calorimetry (DSC) are consistent with the interaction energies calculated at the level of metadynamics. Thus, we present an efficient approach to generate a well-defined DNA-peptide conjugate that mimics a real DNA-peptide complex. These complexes can be used to investigate DNA damage under conditions very similar to those present in the cell.
Assuntos
Fatores de Transcrição de Zíper de Leucina Básica/química , DNA de Cadeia Simples/química , DNA/química , Peptídeos/química , Proteínas de Saccharomyces cerevisiae/química , Sequência de Aminoácidos , Fatores de Transcrição de Zíper de Leucina Básica/metabolismo , Sítios de Ligação , Varredura Diferencial de Calorimetria , Catálise , Cromatografia Líquida de Alta Pressão , Química Click , Cobre/química , DNA/metabolismo , Dano ao DNA , DNA de Cadeia Simples/metabolismo , Simulação de Dinâmica Molecular , Conformação de Ácido Nucleico , Peptídeos/metabolismo , Domínios Proteicos , Proteínas de Saccharomyces cerevisiae/metabolismo , Espectrometria de Massas por Ionização por Electrospray , Temperatura de TransiçãoRESUMO
Basic leucine zipper (bZIP) transcription factors play important roles in numerous growth and developmental processes. Potato (Solanum tuberosum L.) is a worldwide important vegetable crop; nevertheless, no systematic identification or functional analysis of the potato bZIP gene family has been reported. In this research, 65 potato bZIPs distributed on 12 potato chromosomes were identified. According to the topology of Arabidopsis and potato bZIP phylogenetic tree, the bZIPs were classified into thirteen groups, designated as A-K, M, and S, with no potato bZIPs included in groups J and M. The bZIPs from the same group shared a conserved exon-intron structure, intron phase, and motif composition. Eighteen potato bZIPs were involved in segmental duplications, and the duplicated gene pairs were under purifying selection. No tandemly duplicated potato bZIP was found. Each potato bZIP promoter contained at least one kind of stress-responsive or stress-related hormone-responsive element. RNA-seq and qRT-PCR analyses revealed different expression patterns of potato bZIPs under abiotic stresses. The overexpression of StbZIP-65 in Arabidopsis enhanced salt tolerance. The StbZIP-65 protein localized in the nucleus. ß-Glucuronidase staining showed that promoter activity of StbZIP-65 was induced by exogenous methyl jasmonate. These results may aid in further functional studies of potato bZIP transcription factors.
Assuntos
Fatores de Transcrição de Zíper de Leucina Básica/genética , Fatores de Transcrição de Zíper de Leucina Básica/metabolismo , Regulação da Expressão Gênica de Plantas , Tolerância ao Sal , Solanum tuberosum/química , Solanum tuberosum/fisiologia , Arabidopsis/genética , Mapeamento Cromossômico , Duplicação Gênica , Perfilação da Expressão Gênica , Genes de Plantas , Filogenia , Regiões Promotoras Genéticas , Tolerância ao Sal/genética , Solanum tuberosum/classificaçãoRESUMO
Although acetaminophen (APAP) is a commonly used analgesic antipyretic drug, hepatotoxicity and nephrotoxicity are common after the overdose. The main mechanism of APAP toxicity is oxidative stress based. Stress may induce the production of heme oxygenase 1 (HO)-1 which is regulated by interleukin (IL)-10 and inhibit the production of tumor necrosis factor-alpha (TNF-α). HO-1 expression is further regulated by nuclear factor erythroid 2-related factor 2 (Nrf2) and the transcription factor BTB and CNC homology 1 (BACH1). Drug-induced toxicity can be relieved by several natural products, which are preferred due to their dietary nature and less adverse reactions. Of these natural products, omega-3 (ω-3) fatty acids are known for anti-inflammatory and antioxidant actions. However, effects of ω-3fatty acids on APAP-induced hepatic and renal toxicity are not well addressed. We designed this study to test the potential protecting actions of ω-3 fatty acids (270 mg/kg Eicosapentaenoic acid and 180 mg/kg docosahexaenoic acid, orally, for 7 days) in hepatotoxicity and nephrotoxicity induced by APAP (2 g/kg, once orally on day 7) in rats. Moreover, we focused on the molecular mechanism underlying APAP hepatotoxicity and nephrotoxicity. Pre-treatment with ω-3 fatty acids enhanced liver and kidney functions indicated by decreased serum aminotransferases activities and serum creatinine and urea concentrations. These results were further confirmed by histopathological examination. Moreover, ω-3 fatty acids showed antioxidant properties confirmed by decreased malondialdehyde level and increased total antioxidant capacity. Antioxidant Nrf2, its regulators (HO-1 and BACH1) and the anti-inflammatory cytokine (IL-10) were up-regulated by APAP administration as a compensatory mechanism and they were normalized by ω-3 fatty acids. ω-3 fatty acids showed anti-inflammatory actions through down-regulating nuclear factor kappa B (NF-ĸB) and its downstream TNF-α. Moreover, Western blot analysis showed that ω-3 fatty acids promoted Nrf2 translocation to the nucleus; BACH1 exit from the nucleus and inhibited NF-ĸB nuclear translocation. These findings suggested the protecting actions of ω-3 fatty acids against APAP-induced hepatic and renal toxicity through regulation of antioxidant Nrf2 and inflammatory NF-ĸB pathways.
Assuntos
Anti-Inflamatórios/farmacologia , Doença Hepática Induzida por Substâncias e Drogas/prevenção & controle , Ácidos Graxos Ômega-3/farmacologia , Nefropatias/prevenção & controle , Substâncias Protetoras/farmacologia , Transdução de Sinais/efeitos dos fármacos , Acetaminofen , Animais , Fatores de Transcrição de Zíper de Leucina Básica/metabolismo , Núcleo Celular/metabolismo , Doença Hepática Induzida por Substâncias e Drogas/patologia , Regulação para Baixo , Heme Oxigenase (Desciclizante)/metabolismo , Rim/patologia , Nefropatias/induzido quimicamente , Nefropatias/patologia , Fígado/patologia , Masculino , Fator 2 Relacionado a NF-E2/metabolismo , NF-kappa B/metabolismo , Ratos Sprague-Dawley , Proteínas Repressoras/metabolismo , Fator de Crescimento Transformador alfa/metabolismoRESUMO
Abscisic acid (ABA) insensitive 5 (ABI5)-a core transcription factor of the ABA signaling pathway-is a basic leucine zipper transcription factor that plays a key role in the regulation of seed germination and early seedling growth. ABI5 interacts with other phytohormone signals to regulate plant growth and development, and stress responses in Arabidopsis, but little is known about the functions of ABI5 in potatoes. Here, we find that StABI5 is involved in the regulation of chloroplast development and photosynthesis. Genetic analysis indicates that StABI5 overexpression transgenic potato lines accelerate dark-induced leaf yellowing and senescence. The chlorophyll contents of overexpressed StABI5 transgenic potato lines were significantly decreased in comparison to those of wild-type Desiree potatoes under dark conditions. Additionally, the RNA-sequencing (RNA-seq) analysis shows that many metabolic processes are changed in overexpressed StABI5 transgenic potatoes. Most of the genes involved in photosynthesis and carbon fixation are significantly down-regulated, especially the chlorophyll a-b binding protein, photosystem I, and photosystem II. These observations indicate that StABI5 negatively regulates chloroplast development and photosynthesis, and provides some insights into the functions of StABI5 in regard to potato growth.
Assuntos
Fatores de Transcrição de Zíper de Leucina Básica/genética , Fatores de Transcrição de Zíper de Leucina Básica/metabolismo , Cloroplastos/metabolismo , Solanum tuberosum/crescimento & desenvolvimento , Clorofila/metabolismo , Perfilação da Expressão Gênica , Regulação da Expressão Gênica de Plantas , Fotossíntese , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Plantas Geneticamente Modificadas/crescimento & desenvolvimento , Análise de Sequência de RNA , Solanum tuberosum/genética , Solanum tuberosum/metabolismoRESUMO
bZIP transcription factors have been reported to be involved in many different biological processes in plants. The ABA (abscisic acid)-dependent AREB/ABF-SnRK2 pathway has been shown to play a key role in the response to osmotic stress in model plants. In this study, a novel bZIP gene, FtbZIP5, was isolated from tartary buckwheat, and its role in the response to drought and salt stress was characterized by transgenic Arabidopsis. We found that FtbZIP5 has transcriptional activation activity, which is located in the nucleus and specifically binds to ABRE elements. It can be induced by exposure to PEG6000, salt and ABA in tartary buckwheat. The ectopic expression of FtbZIP5 reduced the sensitivity of transgenic plants to drought and high salt levels and reduced the oxidative damage in plants by regulating the antioxidant system at a physiological level. In addition, we found that, under drought and salt stress, the expression levels of several ABA-dependent stress response genes (RD29A, RD29B, RAB18, RD26, RD20 and COR15) in the transgenic plants increased significantly compared with their expression levels in the wild type plants. Ectopic expression of FtbZIP5 in Arabidopsis can partially complement the function of the ABA-insensitive mutant abi5-1 (abscisic acid-insensitive 5-1). Moreover, we screened FtSnRK2.6, which might phosphorylate FtbZIP5, in a yeast two-hybrid experiment. Taken together, these results suggest that FtbZIP5, as a positive regulator, mediates plant tolerance to salt and drought through ABA-dependent signaling pathways.
Assuntos
Fatores de Transcrição de Zíper de Leucina Básica/genética , Proteínas de Plantas/genética , Proteínas Serina-Treonina Quinases/metabolismo , Tolerância ao Sal , Transgenes , Arabidopsis , Fatores de Transcrição de Zíper de Leucina Básica/metabolismo , Fagopyrum/genética , Proteínas de Plantas/metabolismo , Proteínas Serina-Treonina Quinases/genética , TranscriptomaRESUMO
KEY MESSAGE: Overexpression of the tea plant gene CsbZIP18 in Arabidopsis impaired freezing tolerance, and CsbZIP18 is a negative regulator of ABA signaling and cold stress. Basic region/leucine zipper (bZIP) transcription factors play important roles in the abscisic acid (ABA) signaling pathway and abiotic stress response in plants. However, few bZIP transcription factors have been functionally characterized in tea plants (Camellia sinensis). In this study, a bZIP transcription factor, CsbZIP18, was found to be strongly induced by natural cold acclimation, and the expression level of CsbZIP18 was lower in cold-resistant cultivars than in cold-susceptible cultivars. Compared with wild-type (WT) plants, Arabidopsis plants constitutively overexpressing CsbZIP18 exhibited decreased sensitivity to ABA, increased levels of relative electrolyte leakage (REL) and reduced values of maximal quantum efficiency of photosystem II (Fv/Fm) under freezing conditions. The expression of ABA homeostasis- and signal transduction-related genes and abiotic stress-inducible genes, such as RD22, RD26 and RAB18, was suppressed in overexpression lines under freezing conditions. However, there was no significant change in the expression of genes involved in the C-repeat binding factor (CBF)-mediated ABA-independent pathway between WT and CsbZIP18 overexpression plants. These results indicate that CsbZIP18 is a negative regulator of freezing tolerance via an ABA-dependent pathway.
Assuntos
Ácido Abscísico/farmacologia , Arabidopsis/metabolismo , Fatores de Transcrição de Zíper de Leucina Básica/metabolismo , Camellia sinensis/genética , Resposta ao Choque Frio , Congelamento , Regulação da Expressão Gênica de Plantas/efeitos dos fármacos , Aclimatação/genética , Arabidopsis/genética , Proteínas de Arabidopsis/genética , Proteínas de Arabidopsis/metabolismo , Fatores de Transcrição de Zíper de Leucina Básica/genética , Camellia sinensis/metabolismo , Resposta ao Choque Frio/genética , Regulação da Expressão Gênica de Plantas/genética , Complexo de Proteína do Fotossistema II/metabolismo , Filogenia , Plantas Geneticamente Modificadas/genética , Plantas Geneticamente Modificadas/metabolismo , Proteostase/efeitos dos fármacos , Proteostase/genética , Transdução de Sinais/efeitos dos fármacos , Transdução de Sinais/genética , Fatores de Transcrição/genética , Fatores de Transcrição/metabolismo , Regulação para Cima , Proteínas rab de Ligação ao GTP/genética , Proteínas rab de Ligação ao GTP/metabolismoRESUMO
The basic region-leucine zipper (bZIP) transcription factors (TFs) form homodimers and heterodimers via the coil-coil region. The bZIP dimerization network influences gene expression across plant development and in response to a range of environmental stresses. The recent release of the most comprehensive potato reference genome was used to identify 80 StbZIP genes and to characterize their gene structure, phylogenetic relationships, and gene expression profiles. The StbZIP genes have undergone 22 segmental and one tandem duplication events. Ka/Ks analysis suggested that most duplications experienced purifying selection. Amino acid sequence alignments and phylogenetic comparisons made with the Arabidopsis bZIP family were used to assign the StbZIP genes to functional groups based on the Arabidopsis orthologs. The patterns of introns and exons were conserved within the assigned functional groups which are supportive of the phylogeny and evidence of a common progenitor. Inspection of the leucine repeat heptads within the bZIP domains identified a pattern of attractive pairs favoring homodimerization, and repulsive pairs favoring heterodimerization. These patterns of attractive and repulsive heptads were similar within each functional group for Arabidopsis and S. tuberosum orthologs. High-throughput RNA-seq data indicated the most highly expressed and repressed genes that might play significant roles in tissue growth and development, abiotic stress response, and response to pathogens including Potato virus X. These data provide useful information for further functional analysis of the StbZIP gene family and their potential applications in crop improvement.
Assuntos
Fatores de Transcrição de Zíper de Leucina Básica/genética , Família Multigênica , Solanum tuberosum/genética , Motivos de Aminoácidos , Fatores de Transcrição de Zíper de Leucina Básica/metabolismo , Sítios de Ligação , Mapeamento Cromossômico , Sequência Conservada , Éxons , Perfilação da Expressão Gênica , Regulação da Expressão Gênica de Plantas/efeitos dos fármacos , Ordem dos Genes , Genoma de Planta , Íntrons , Especificidade de Órgãos , Filogenia , Regiões Promotoras Genéticas , Solanum tuberosum/classificação , Solanum tuberosum/metabolismo , Estresse Fisiológico , TranscriptomaRESUMO
Aging is a degenerative process characterized by progressive deterioration of cellular components, ultimately resulting in mortality, in which massive accumulation of reactive oxygen species (ROS) and advanced glycation end products (AGEs) are implicated as crucial factors. At the same time, natural products are rich sources from which to isolate and characterize potential anti-aging compounds. The current study was designed to extract compounds from the marine bacterium Pseudomonas sp. and investigate their in vitro antioxidant and anti-glycation activities, as well as their in vivo effects on aging in the model organism Schizosaccharomyces pombe. In vitro assays showed that a Pseudomonas sp. PTR-08 extract exhibited the best antioxidant and anti-glycation activities. Further, direct administration of the extract significantly increased yeast longevity, accompanied by induction of the yeast oxidative stress response. Molecular analyses indicated that selected extract dramatically up-regulated the expression of pap1+, which encodes the transcriptional factor Pap1 and ctt1+, which encodes catalase, following H2O2 treatment. In line with these results, catalase activity significantly increased, leading to a decrease in intracellular ROS. In addition, this extract may delay the G1 phase of the yeast cell cycle, leading to an extended lifespan. Moreover, our findings indicated that the extract contains pyrrolo[1,2-a]pyrazine-1,4-dione, hexahydro-, which substantially promotes anti-aging activity in yeast. However, further research must be conducted to better understand the role of this compound in our system.
Assuntos
Antioxidantes/isolamento & purificação , Antioxidantes/farmacologia , Ciclo Celular/efeitos dos fármacos , Senescência Celular/efeitos dos fármacos , Pseudomonas/química , Schizosaccharomyces/efeitos dos fármacos , Organismos Aquáticos , Fatores de Transcrição de Zíper de Leucina Básica/genética , Fatores de Transcrição de Zíper de Leucina Básica/metabolismo , Catalase/genética , Catalase/metabolismo , Ciclo Celular/genética , Avaliação Pré-Clínica de Medicamentos , Regulação Fúngica da Expressão Gênica/efeitos dos fármacos , Longevidade/efeitos dos fármacos , Longevidade/genética , Organismos Geneticamente Modificados , Estresse Oxidativo/efeitos dos fármacos , Estresse Oxidativo/genética , Espécies Reativas de Oxigênio/metabolismo , Schizosaccharomyces/citologia , Schizosaccharomyces/fisiologia , Proteínas de Schizosaccharomyces pombe/genética , Proteínas de Schizosaccharomyces pombe/metabolismo , Transdução de Sinais/efeitos dos fármacos , Transdução de Sinais/genéticaRESUMO
ETHNOPHARMACOLOGICAL RELEVANCE: San Wei Gan jiang San (SWGJS) also called Jia Ga Song Tang, is widely used in ancient medicine for liver diseases. THE AIM OF THIS STUDY: To identify the blood components of SWGJS. To determine the hepatoprotective effect and the mechanism of SWGJS by observing its effect on different degrees of liver damage and gene knockdown cells. MATERIALS AND METHODS: SWGJS treated serum was analyzed by UPLC-MS to identify blood components. CCl4-induced chronic liver injury in rats was treated with SWGJS. The viscera index was calculated. Pathological changes of the liver were determined by HE staining and analysis of by following: GSH-Px and MDA in liver homogenate; ALT and AST in serum; mRNA expression of Nrf2, Bach1, and HO-1 by RT-PCR; Nrf2 and Bach1 in the nucleus and cytoplasm; HO-1 total expression by Western blot; silencing Nrf2 and Bach1 in human L-02 cells by siRNA; MDA, GSH-Px, GST, and GR in cell supernatants; and GSH/GSSG within the cell. RESULTS: We found that 6-gingerol was one of the blood components in the serum treated with SWGJS. In CCl4-induced chronic liver injury in rats, SWGJS repaired the liver structure in the early stages of liver damage as evidenced by reduced ALT and AST in the serum, increased GSH-Px activity and decreased MDA levels in the liver over time. SWGJS has excellent antioxidant and hepatoprotective effects and prevents disease progression. The mechanism of SWGJS is related to the dynamics promoting Nrf2 entry to the nucleus and Bach1 exit from the nucleus. In L-02 cells with silenced Nrf2, the antioxidant enzyme system was disordered, and the change in the cellular redox state was not conducive to antioxidative stress. However, in cells with silenced Bach1, the antioxidant enzyme system could be activated to promote cellular antioxidant stress. SWGJS had a combined effect on Nrf2 and Bach1 contributing to antioxidant properties and liver protection. SWGJS increased GSH-Px and HO-1, decreased MDA and increased the ratio of GSH/GSSG by upregulating the expression of Nrf2 to enhance its antioxidant effects. At the same time, SWGJS had a specific impact on decreasing Bach1. Its elevation of GST is due to the overall performance of increasing Nrf2 and decreasing Bach1. This mechanism of action embodies the characteristics of the multitarget impact of traditional medicine and the antioxidation effect of SWGJS. CONCLUSIONS: 6-Gingerol is one of the blood components of SWGJS. SWGJS can regulate antioxidant enzymes, protect against liver damage in different stages, and slow the progression of liver cell damage and liver disease by increasing Nrf2 and reducing Bach1 in the nucleus, dynamically regulating Nrf2/Bach1.
Assuntos
Doença Hepática Induzida por Substâncias e Drogas/tratamento farmacológico , Animais , Fatores de Transcrição de Zíper de Leucina Básica/genética , Fatores de Transcrição de Zíper de Leucina Básica/metabolismo , Tetracloreto de Carbono , Doença Hepática Induzida por Substâncias e Drogas/genética , Doença Hepática Induzida por Substâncias e Drogas/metabolismo , Doença Hepática Induzida por Substâncias e Drogas/patologia , Heme Oxigenase (Desciclizante)/genética , Heme Oxigenase (Desciclizante)/metabolismo , Fígado/efeitos dos fármacos , Fígado/metabolismo , Fígado/patologia , Masculino , Fator 2 Relacionado a NF-E2/genética , Fator 2 Relacionado a NF-E2/metabolismo , Ratos Sprague-DawleyRESUMO
Astragaloside â £(AS-â £) has protective effects against ischemia-reperfusion injury(IRI), but its mechanism of action has not yet been determined. This study aims to investigate the protective effects and mechanism of AS-â £ on H9c2 cardiomyocyte injury induced by hypoxia-reoxygenation(H/R). The H/R model of myocardial cells was established by hypoxic culture for 12 hours and then reoxygenation culture for 8 hours. After AS-â £ treatment, cell viability, the reactive oxygen species(ROS) levels, as well as the content or activity of superoxide dismutase(SOD), malondialdehyde(MDA), interleukin 6(IL-6), and tumor necrosis factor alpha(TNF-α), were measured to evaluate the effect of AS-â £ treatment. The effect of AS-â £ on HO-1 protein expression and nuclear Nrf2 and Bach1 protein expression was determined by Western blot. Finally, siRNA was used to knock down HO-1 gene expression to observe its reversal effect on AS-â £ intervention. The results showed that as compared with the H/R model group, the cell viability was significantly increased(P<0.01), ROS level in the cells, MDA, hs-CRP and TNF-α in cell supernatant and nuclear protein Bach1 expression in the cells were significantly decreased(P<0.01), while SOD content, HO-1 protein expression in cells and expression of nuclear protein Nrf2 were significantly increased(P<0.01) in H/R+AS-â £ group. However, pre-transfection of HO-1 siRNA into H9c2 cells by liposome could partly reverse the above effects of AS-â £ after knocking down the expression of HO-1. This study suggests that AS-â £ has significant protective effect on H/R injury of H9c2 cardiomyocytes, and Nrf2/Bach1/HO-1 signaling pathway may be a key signaling pathway for the effect.