Promoter Elements Research Articles

Genome-wide identification and expression analysis of the GASA gene family in Chinese cabbage (Brassica rapa L. ssp. pekinensis)

BackgroundThe Gibberellic Acid-Stimulated Arabidopsis (GASA) gene family is widely involved in the regulation of plant growth, development, and stress response. However, information on the GASA gene family has not been reported in Chinese cabbage (Brassica rapa L. ssp. pekinensis).ResultsHere, we conducted genome-wide identification and analysis of the GASA genes in Chinese cabbage. In total, 15 GASA genes were identified in the Chinese cabbage genome, and the physicochemical property, subcellular location, and tertiary structure of the corresponding GASA proteins were elucidated. Phylogenetic analysis, conserved motif, and gene structure showed that the GASA proteins were divided into three well-conserved subfamilies. Synteny analysis proposed that the expansion of the GASA genes was influenced mainly by whole-genome duplication (WGD) and transposed duplication (TRD) and that duplication gene pairs were under negative selection. Cis-acting elements of the GASA promoters were involved in plant development, hormonal and stress responses. Expression profile analysis showed that the GASA genes were widely expressed in different tissues of Chinese cabbage, but their expression patterns appeared to diverse. The qRT-PCR analysis of nine GASA genes confirmed that they responded to salt stress, heat stress, and hormonal triggers.ConclusionsOverall, this study provides a theoretical basis for further exploring the important role of the GASA gene family in the functional genome of Chinese cabbage.

The bipartite promoter of Orthonairovirus hazaraense large segment.

The realization that segmented negative-strand RNA virus genome ribonucleoproteins are never free as their RNA ends are always bound to the viral polymerase has highlighted the problem of how these genome segments are replicated and express their mRNAs while their RNA ends remain associated with the polymerase throughout the cycles of RNA synthesis. This study of the length and nucleotide composition of the Orthonairovirus hazaraense L segment-specific double-stranded RNA (dsRNA) promoter element (the promoter duplex) provides insight into how its mRNA might be initiated and suggests that this promoter element acts via its separated single strands as well as via dsRNA.

Circuit-specific gene therapy reverses core symptoms in a primate Parkinson’s disease model

Parkinson's disease (PD) is a debilitating neurodegenerative disorder. Its symptoms are typically treated with levodopa or dopamine receptor agonists, but its action lacks specificity due to the wide distribution of dopamine receptors in the central nervous system and periphery. Here, we report the development of a gene therapy strategy to selectively manipulate PD-affected circuitry. Targeting striatal D1 medium spiny neurons (MSNs), whose activity is chronically suppressed in PD, we engineered a therapeutic strategy comprised of a highly efficient retrograde adeno-associated virus (AAV), promoter elements with strong D1-MSN activity, and a chemogenetic effector to enable precise D1-MSN activation after systemic ligand administration. Application of this therapeutic approach rescues locomotion, tremor, and motor skill defects in both mouse and primate models of PD, supporting the feasibility of targeted circuit modulation tools for the treatment of PD in humans.

Identification and expression analysis of the KNOX genes during organogenesis and stress responseness in Camellia sinensis (L.) O. Kuntze.

Tea plant (Camellia sinensis L.), whose leaves are the major reproductive organs, has been cultivated and consumed widely for its economic and health benefits. The Knotted1-like Homeobox (KNOX) proteins play significant roles in leaf morphology formation and development. However, the functions of KNOX proteins in tea plants are still unknown. Here, 11 CsKNOX genes from the tea plants were cloned and divided into Class I, II, and KNATM clades based on their protein sequences. These 11 CsKNOX genes were mapped on 8 out of 15 tea plant chromosomes, all localized in the nucleus. Specific spatiotemporal expression patterns of CsKNOX genes were found in various tissues and different development periods of buds, flowers, and roots of tea plants. Meanwhile, transcript levels of CsKNOX in tea leaves were strongly correlated with the accumulation of flavan-3-ols and proanthocyanidins. It was found that most of the CsKNOX genes could respond to drought, salt, cold, and exogenous MeJA and GA3 by analysis of transcriptomics data and promoter elements. The protein interaction analysis showed that CsKNOX could cooperate with CsAS1 and other critical functional proteins. In conclusion, this research provided the basic information for the functions of the CsKNOX family during organogenesis and stress response in tea plants, which was necessary for further functional characterization verification.

Pengembangan dan Implementsi Strategi Pemasaran Inovatif untuk Produk Rengginang Mini dalam Industri Makanan Ringan Metode Analisis SWOT dan Marketing MIX

Mini rengginang is a traditional Indonesian food which has been innovated into a snack and can be consumed by all groups, both young and old. This research aims to identify and implement innovative marketing strategies to increase market penetration of mini rengginang products in the snack food industry. The SWOT evaluation method is used to analyze the strengths, weaknesses, opportunities and threats associated with a product in a competitive market. Meanwhile, the application of the marketing mix aims to optimize product, price, promotion and distribution elements in order to achieve the specified marketing objectives. By integrating these two methods, this research seeks to provide a comprehensive view of how to address market challenges, identify competitive advantages, and adapt marketing strategies to gain a competitive edge in the ever-evolving snack food industry.

Overexpressing CsPSY1 Gene of Tea Plant, Encoding a Phytoene Synthase, Improves α-Carotene and β-Carotene Contents in Carrot.

Tea plants (Camellia sinensis (L.) O. Kuntze) belong to Theaceae family, in the section Thea. Tea plants are widely distributed in subtropical and tropical regions in the word. α-carotene and β-carotene in the tea leaves belong to carotenoids, which are associated with the aroma and color of the tea. Phytoene synthase (PSY) is a rate-limiting enzyme in carotenoids biosynthesis. We identified three CsPSY genes in 'Shuchazao', named CsPSY1, CsPSY2, and CsPSY3. Structural analysis of three CsPSY genes showed that CsPSY1 had a longer intro structure. The cis-acting elements of CsPSYs promoter were mainly associated with light-responsiveness, abiotic stress-responsiveness, and hormone-responsiveness. CsPSY1 exhibited expression in all tissues of the tea plants, whereas CsPSY2 and CsPSY3 were trace expression levels in all tissues. The positive expression of CsPSY1 under hormonal and abiotic stresses suggested its role in plant development and defense responses. The amino acid sequence of CsPSY1 was highly conserved in eight tea cultivars. The recombinant vector pCAMBIA1301-CsPSY1 was constructed to stabilize the overexpression of CsPSY1 in carrot. The contents of α-carotene and β-carotene in transgenic carrot callus were significantly increased. This study provides a foundational basis for further research on the function of CsPSYs and carotenoids accumulation in tea plants.

Understanding the success of a deaf-funder’s restaurant between food and empathy’s appeal: The case of 1001 signes

Deaf from birth and of Moroccan origin, Sid Mouar has been a successful restaurant owner for 8 years. His restaurant specializes in serving Moroccan cuisine and employs individuals with disabilities. The ordering process utilizes sign language for communication. This case study delves into the reasons behind the restaurant’s success by examining the story of the deaf entrepreneur with a disability and by identifying all the service attributes mentioned in customer feedback. By utilizing the tetraclass model, the impact of each service attribute on customer satisfaction and dissatisfaction is evaluated. This analysis offers insights into what aspects are fundamental, crucial, additional, or secondary in terms of customer satisfaction, thus contributing to an understanding of the restaurant’s achievement. The case also delves into topics related to service quality, customer satisfaction, competitive advantages, promotional elements, and the restaurant’s future adaptations to remain competitive.

A comparative study on the influence of potential 4th period promoters in Ni-bimetallic catalysts for CO2 methanation

Because of their simplicity, bimetallic nickel catalysts are a rational step to further develop improved CO2-methanation catalysts suitable for specific applications and feed gas situations. In this study, hydrotalcite-based catalysts incorporating a second metal from the 4th period (chromium, manganese, iron, cobalt, copper, zinc and gallium) were synthesized, characterized (ICP-OES, XRD, BET, XPS) and performance tested under comparable conditions. The study aims at the clarification of inconsistencies regarding these additives found in the literature by ensuring comparability as good as possible. To achieve this goal the catalysts were tested using the same reactor, were prepared by the same type of synthesis possessing the same atomic ratio of Ni to the potential promoter element. In catalytic tests, performed at atmospheric pressure and at 20 bar, all catalysts were active in respect to CO2 hydrogenation. While Cu, Zn, and Ga acted as inhibitors, reducing the activity approximately by half, Fe and Mn proved to be very active promotors. The catalyst without promotor shows 80 % yield at 400 °C whereas the iron containing catalyst reaches this value already at 320 °C, but undergoes a irreversible loss at high temperatures (20 % loss after being exposed to 480 °C). In contrast, the manganese containing catalyst possesses a significantly improved activity without showing such a deactivation process, indicating a high level of stability.

Repression of GhTUBB1 Reduces Plant Height in Gossypium hirsutum.

The original 'Green Revolution' genes are associated with gibberellin deficiency. However, in some species, mutations in these genes cause pleiotropic phenotypes, preventing their application in dwarf breeding. The development of novel genotypes with reduced plant height will resolve this problem. In a previous study, we obtained two dwarf lines, L28 and L30, by introducing the Ammopiptanthus mongolicus (Maxim. ex Kom.) Cheng f. C-repeat-binding factor 1 (AmCBF1) into the upland cotton variety R15. We found that Gossypium hirsutum Tubulin beta-1 (GhTUBB1) was downregulated in L28 and L30, which suggested that this gene may have contributed to the dwarf phenotype of L28 and L30. Here, we tested this hypothesis by silencing GhTUBB1 expression in R15 and found that decreased expression resulted in a dwarf phenotype. Interestingly, we found that repressing AmCBF1 expression in L28 and L30 partly recovered the expression of GhTUBB1. Thus, AmCBF1 expression presented a negative relationship with GhTUBB1 expression in L28 and L30. Moreover, yeast one-hybrid and dual-luciferase assays suggest that AmCBF1 negatively regulates GhTUBB1 expression by directly binding to C-repeat/dehydration-responsive (CRT/DRE) elements in the GhTUBB1 promoter, potentially explaining the dwarf phenotypes of L28 and L30. This study elucidates the regulation of GhTUBB1 expression by AmCBF1 and suggests that GhTUBB1 may be a new target gene for breeding dwarf and compact cultivars.

The Arabidopsis SHORTROOT network coordinates shoot apical meristem development with auxin-dependent lateral organ initiation.

Plants produce new organs post-embryonically throughout their entire life cycle. This is due to stem cells present in the shoot and root apical meristems, the SAM and RAM, respectively. In the SAM, stem cells are located in the central zone where they divide slowly. Stem cell daughters are displaced laterally and enter the peripheral zone, where their mitotic activity increases and lateral organ primordia are formed. How the spatial arrangement of these different domains is initiated and controlled during SAM growth and development, and how sites of lateral organ primordia are determined in the peripheral zone is not yet completely understood. We found that the SHORTROOT (SHR) transcription factor together with its target transcription factors SCARECROW (SCR), SCARECROW-LIKE23 (SCL23) and JACKDAW (JKD), promotes formation of lateral organs and controls shoot meristem size. SHR, SCR, SCL23, and JKD are expressed in distinct, but partially overlapping patterns in the SAM. They can physically interact and activate expression of key cell cycle regulators such as CYCLIND6;1 (CYCD6;1) to promote the formation of new cell layers. In the peripheral zone, auxin accumulates at sites of lateral organ primordia initiation and activates SHR expression via the auxin response factor MONOPTEROS (MP) and auxin response elements in the SHR promoter. In the central zone, the SHR-target SCL23 physically interacts with the key stem cell regulator WUSCHEL (WUS) to promote stem cell fate. Both SCL23 and WUS expression are subject to negative feedback regulation from stem cells through the CLAVATA signaling pathway. Together, our findings illustrate how SHR-dependent transcription factor complexes act in different domains of the shoot meristem to mediate cell division and auxin dependent organ initiation in the peripheral zone, and coordinate this activity with stem cell maintenance in the central zone of the SAM.

Validating a Promoter Library for Application in Plasmid-Based Diatom Genetic Engineering.

While diatoms are promising synthetic biology platforms, there currently exists a limited number of validated genetic regulatory parts available for genetic engineering. The standard method for diatom transformation, nonspecific introduction of DNA into chromosomes via biolistic particle bombardment, is low throughput and suffers from clonal variability and epigenetic effects. Recent developments in diatom engineering have demonstrated that autonomously replicating episomal plasmids serve as stable expression platforms for diverse gene expression technologies. These plasmids are delivered via bacterial conjugation and, when combined with modular DNA assembly technologies, provide a flexibility and speed not possible with biolistic-mediated strain generation. In order to expand the current toolbox for plasmid-based engineering in the diatom Phaeodactylum tricornutum, a conjugation-based forward genetics screen for promoter discovery was developed, and application to a diatom genomic DNA library defined 252 P. tricornutum promoter elements. From this library, 40 promoter/terminator pairs were delivered via conjugation on episomal plasmids, characterized in vivo, and ranked across 4 orders of magnitude difference in reporter gene expression levels.

Adaptation and psychometric properties of the Greek version of Youth Efficacy/Empowerment Scale – Mental Health

Empowerment as a crucial element of health promotion has increasingly been prioritized among people who are capable of raising their voice and having a meaningful input in their care. The “Youth Efficacy/Empowerment Scale-Mental health” (YES-MH) is a measure designed to evaluate empowerment from the perspective of young service users living with mental health problems. The present study aimed to examine the psychometric properties of the YES-MH among youth clinical population in Greece and secondarily to analyze the relationship between empowerment and socio-demographic variables. Data were collected in mental health care settings of Athens from 150 participants aged 12 to 17 years. Exploratory factor analysis was conducted by the method of Principal Components to determine the structure. Validity was tested by comparing the YES-MH with two scales and reliability by using Cronbach alpha coefficient. Results indicated the predicted three-factor solution and acceptable internal consistency (0.76 to 0.86). Concerning the concurrent validity, YES-MH significantly correlated with mental well-being (r = 0.60) and the presence of difficulties (r = −0.34)”. Moreover, empowerment was associated to gender and family economic status at the self-level and significantly higher scores were observed to the younger. The current study added evidence for the validity and reliability of the Greek version. Its implications extend from research to practice including the evaluation of interventions and the improvement of services.

Genome-wide characterization of tobacco B-BOX gene family identified two close members play contrast roles under cold stress

Cold stress influences the growth and geographical distribution of plants. Tobacco (Nicotiana tabacum L.), a model in plant biology studies and a world-widely cultivated economic plant from tropical America, exhibits extreme sensitivity to low temperatures. The conserved B-BOX (BBX) transcription factors play vital roles in plant photomorphogenesis and flowering regulation, and their functions in abiotic stresses, including cold stress, have been uncovered recently. However, few studies on tobacco BBX genes have been conducted to date. In this study, 49 non-redundant BBX genes have been identified in the reference tobacco genome. Gene structure, phylogenetic relationship, prediction of cis-regulatory elements in promoters, tissue-specific expression patterns, and response to cold stress were conducted on these BBX gene members. The expression of NtBBX9 and NtBBX11, two close members in the amplified group II, was up-regulated under cold treatments. Overexpression of NtBBX11 embedded the cold tolerance of transgenic plants, while the NtBBX9 overexpressors were susceptible to cold. The NtBBX9 functions as a negative regulator of tobacco response to cold stress, which was confirmed by the phenotype of NtBBX9 RNA-silenced lines. Furthermore, we demonstrated that NtBBX9 and NtBBX11 regulated the cold stress response by alternating the expression of critical cold-regulated genes such as CBFs, LEA14, and LTI65 and reactive oxygen species (ROS) production. The combination of NtBBX9 and NtBBX11 could facilitate tobacco’s rapid response to and recovery from transient cold stress and regulate plant cold-responsive growth in tropical regions.

The KIR2DL1 intermediate upstream element participates in gene activation

The human KIR genes encode a family of class I MHC receptors that are expressed on subsets of NK cells. The expression of KIR proteins is controlled by a stochastic process, and competition between sense and antisense promoter elements has been suggested to program the variegated expression of these genes. Previous studies have demonstrated distinct roles of distal, intermediate, and proximal sense promoter/enhancer elements in gene activation and expression. Conversely, proximal and intronic antisense promoter transcripts have been associated with gene silencing at different stages of NK cell development. In the current study, we examine the effect of intermediate promoter deletion on KIR2DL1 expression in the YTS cell line. Homozygous deletion of the KIR2DL1 intermediate element did not affect proximal promoter activity but resulted in increased detection of upstream transcripts. No significant changes in alternative mRNA splicing or expression levels of KIR2DL1 protein were observed. However, intermediate element deletion was associated with a reduced frequency of gene activation by 5-azacytidine. Taken together, these results indicate that the intermediate element is not an enhancer required for KIR expression; however, it is required for the efficient activation of the gene.

Differential adenine methylation analysis reveals increased variability in 6mA in the absence of methyl-directed mismatch repair.

Methylation greatly influences the bacterial genome by guiding DNA repair and regulating pathogenic and stress-response phenotypes. But, the rate of epigenetic changes and their consequences on molecular phenotypes are underexplored. Through a detailed characterization of genome-wide adenine methylation in a commonly used laboratory strain of Escherichia coli, we reveal that mismatch repair deficient populations experience an increase in epimutations resulting in a genome-wide reduction of 6mA methylation in a manner consistent with genetic drift. Our findings highlight how methylation patterns evolve and the constraints on epigenetic evolution due to post-replicative DNA repair, contributing to a deeper understanding of bacterial genome evolution and how epimutations may introduce semi-permanent variation that can influence adaptation.

The Adr1 transcription factor directs regulation of the ergosterol pathway and azole resistance in Candida albicans.

Research often relies on well-studied orthologs within related species, with researchers using a well-studied gene or protein to allow prediction of the function of the ortholog. In the opportunistic pathogen Candida albicans, orthologs are usually compared with Saccharomyces cerevisiae, and this approach has been very fruitful. Many transcription factors (TFs) do similar jobs in the two species, but many do not, and typically changes in function are driven not by modifications in the structures of the TFs themselves but in the connections between the transcription factors and their regulated genes. This strategy of changing TF function has been termed transcription factor rewiring. In this study, we specifically looked for rewired transcription factors, or Candida-specific TFs, that might play a role in drug resistance. We investigated 30 transcription factors that were potentially rewired or were specific to the Candida clade. We found that the Adr1 transcription factor conferred resistance to drugs like fluconazole, amphotericin B, and terbinafine when activated. Adr1 is known for fatty acid and glycerol utilization in Saccharomyces, but our study reveals that it has been rewired and is connected to ergosterol biosynthesis in Candida albicans.

Logotype as a Tool in Marketing Communication: An Analysisof Websites of Agritourism Farms

A logotype can have a considerable impact on the customers’ perception of a given business, generate experiences and become a basis for assigning specific associations to the brand (positive, negative, promotional). The aim of the present study is to verify the function of logotypes of selected agritourism establishments, especially in view of the rationale for placing them on home pages of websites, and the marketing function they fulfill. Homepages (the main web pages) of websites of 10 agritourism farms with the dimensions of 1,245 × 570 pixels (and 24 -bit color depth) were analyzed. The study involved 170 respondents ( n = 170), whose task was to indicate a single element they perceive as positive, negative, and one that in their opinion best serves the promotional goal in each of the 10 homepages. Statistical analyses were performed using the R-CRAN software version 4.0.2. The study confirmed that there are both differences and similarities in the responses regarding logotypes of the agritourism farms studied. The perception of the logotype depends strongly on the education level and age of respondents. The analyses also confirmed that there is no linear relationship between the size of the logo and the frequency at which it is indicated as a promotional element. The use of a detailed factor analysis of logotypes (with the use of the click-tracking technique) on the websites of agritourism farms is an innovative approach and allows for a relatively comprehensive assessment of the perception of logotypes by potential consumers of offers available on the market.

An ABA-inducible gene IbTSJT1 positively regulates drought tolerance in transgenic sweetpotato

The TSJT1 protein belongs to the superfamily of class-II glutamine amidotransferases (GATase). There is little research on the functions and underlying mechanisms of TSJT1 in plants. In this study, an abscisic acid (ABA)-inducible gene IbTSJT1 was isolated from drought-tolerant sweetpotato line ‘Xushu55-2’. Its expression was strongly induced by PEG6000 and ABA. The IbTSJT1 protein was localized in the nucleus and cell membrane. The IbTSJT1-overexpressing sweetpotato plants exhibited significantly enhanced drought tolerance. Their ABA and proline contents and superoxide dismutase (SOD) and peroxidase (POD) activities were increased and reactive oxygen species (ROS) scavenging-related genes were upregulated under drought stress. The stomatal aperture assay confirmed that the IbTSJT1-overexpressing plants increased the sensitivity to ABA. Yeast one-hybrid (Y1H) assay, electrophoretic mobility shift assay (EMSA), luciferase reporter assay and ChIP-qPCR assay indicated that IbABF2 could directly bind to the cis-acting ABA-responsive element (ABRE) in the IbTSJT1 promoter to activate the expression of IbTSJT1. These findings suggest that IbTSJT1 mediates ABA-dependent drought stress responses and enhances drought tolerance by inducing stomatal closure and activating ROS scavenging system in transgenic sweetpotato. Our study provides a novel gene for improving drought tolerance of sweetpotato and other plants.

Genome-wide identification of bZIP family genes and screening of candidate AarbZIPs involved in terpenoid biosynthesis in Artemisia argyi

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.

Targeting cis-regulatory elements of FOXO family is a novel therapeutic strategy for induction of leukemia cell differentiation

Differentiation therapy has been proposed as a promising therapeutic strategy for acute myeloid leukemia (AML); thus, the development of more versatile methodologies that are applicable to a wide range of AML subtypes is desired. Although the FOXOs transcription factor represents a promising drug target for differentiation therapy, the efficacy of FOXO inhibitors is limited in vivo. Here, we show that pharmacological inhibition of a common cis-regulatory element of forkhead box O (FOXO) family members successfully induced cell differentiation in various AML cell lines. Through gene expression profiling and differentiation marker-based CRISPR/Cas9 screening, we identified TRIB1, a complement of the COP1 ubiquitin ligase complex, as a functional FOXO downstream gene maintaining an undifferentiated status. TRIB1 is direct target of FOXO3 and the FOXO-binding cis-regulatory element in the TRIB1 promoter, referred to as the FOXO-responsive element in the TRIB1 promoter (FRE-T), played a critical role in differentiation blockade. Thus, we designed a DNA-binding pharmacological inhibitor of the FOXO-FRE-T interface using pyrrole-imidazole polyamides (PIPs) that specifically bind to FRE-T (FRE-PIPs). The FRE-PIPs conjugated to chlorambucil (FRE-chb) inhibited transcription of TRIB1, causing differentiation in various AML cell lines. FRE-chb suppressed the formation of colonies derived from AML cell lines but not from normal counterparts. Administration of FRE-chb inhibited tumor progression in vivo without remarkable adverse effects. In conclusion, targeting cis-regulatory elements of the FOXO family is a promising therapeutic strategy that induces AML cell differentiation.