ABSTRACT
Organic-inorganic hybrid low-dimensional lead halides have garnered significant interest in the realm of solid-state optical materials due to their unique properties and potential applications. In this study, we report the synthesis, characterization and application of Mn2+-doped one-dimensional (1D) [AEP]PbCl5·H2O hybrid lead halide perovskites with tunable photoluminescence properties. The Mn2+ doping leads to a redshift of the dominant emission wavelength from 463 nm to 630 nm, with the optimal doping concentration resulting in an enhanced photoluminescence quantum yield (PLQY) from less than 1% to 8.96%. The structural and optical stability of these doped perovskites have been thoroughly investigated revealing excellent performance under humid and high-temperature conditions. Perovskite-PVP composite films exhibit high crystallization and bright orange-red emission under UV excitation. Furthermore, we demonstrate the successful fabrication of a white LED device using the Mn2+-doped perovskite in combination with commercial green and blue phosphors. The fabricated LED exhibits a high color rendering index (CRI) of 87.2 and stable electroluminescence performance under various operating currents and extended operation times. Our findings highlight the potential of Mn2+-doped 1D hybrid lead halide perovskites as efficient and stable phosphors for high-performance white light emitting diodes and other optoelectronic applications.
ABSTRACT
Glucose can serve as both the source of energy and regulatory signaling molecule in plant. Due to the environmental and metabolic change, sugar levels could affect various developmental processes. High glucose environment is hardly conductive to the plant growth but cause development arrest. Increasing evidence indicate that alternative splicing (AS) plays a pivotal role in sugar signaling. However, the regulatory mechanism upon glucose stress remains unclear. The full-length transcriptomes were obtained from the samples of Arabidopsis seedlings with 3% glucose and mock treatment, using Oxford Nanopore sequencing technologies. Further analysis indicated that many genes involved in photosynthesis were significantly repressed and many genes involved in glycolysis, mitochondrial function, and the response to oxidative stress were activated. In total, 1,220 significantly differential alternative splicing (DAS) events related to 619 genes were identified, among which 75.74% belong to intron retention (IR). Notably, more than 20% of DAS events come from a large set of glycine-rich protein (GRP) family genes, such as GRP7, whose AS types mostly belong to IR. Besides the known productive GRP transcript isoforms, we identified a lot of splicing variants with diverse introns spliced in messenger RNA (mRNA) region coding the glycine-rich (GR) domain. The AS pattern of GRPs changed and particularly, the productive GRPs increased upon glucose stress. These ASs of GRP pre-mRNAs triggered by glucose stress could be abolished by AZD-8055, which is an ATP competitive inhibitor for the target of rapamycin (TOR) kinase but could be mimicked by H2O2. Additionally, AS pattern change of arginine/serine-rich splicing factor 31(RS31) via TOR pathway, which was previously described in response to light and sucrose signaling, was also induced in a similar manner by both glucose stress and reactive oxygen species (ROS). Here we conclude that (i) glucose stress suppresses photosynthesis and activates the glycolysis-mitochondria energy relay and ROS scavenging system; (ii) glucose stress triggers transcriptome-wide AS pattern changes including a large set of splicing factors, such as GRPs and RS31; (iii) high sugars regulate AS pattern change of both GRPs and RS31 via TOR-ROS pathway. The results from this study will deepen our understanding of the AS regulation mechanism in sugar signaling.
ABSTRACT
Communication between interacting organisms via bioactive molecules is widespread in nature and plays key roles in diverse biological processes. Small RNAs (sRNAs) can travel between host plants and filamentous pathogens to trigger transkingdom RNA interference (RNAi) in recipient cells and modulate plant defense and pathogen virulence. However, how fungal pathogens counteract transkingdom antifungal RNAi has rarely been reported. Here we show that a secretory protein VdSSR1 (secretory silencing repressor 1) from Verticillium dahliae, a soil-borne phytopathogenic fungus that causes wilt diseases in a wide range of plant hosts, is required for fungal virulence in plants. VdSSR1 can translocate to plant nucleus and serve as a general suppressor of sRNA nucleocytoplasmic shuttling. We further reveal that VdSSR1 sequesters ALY family proteins, adaptors of the TREX complex, to interfere with nuclear export of the AGO1microRNA (AGO1miRNA) complex, leading to a great attenuation in cytoplasmic AGO1 protein and sRNA levels. With this mechanism, V. dahliae can suppress the accumulation of mobile plant miRNAs in fungal cells and succedent transkingdom silencing of virulence genes, thereby increasing its virulence in plants. Our findings reveal a mechanism by which phytopathogenic fungi antagonize antifungal RNAi-dependent plant immunity and expand the understanding on the complex interaction between host and filamentous pathogens.
Subject(s)
MicroRNAs , Verticillium , Active Transport, Cell Nucleus , Antifungal Agents , MicroRNAs/genetics , MicroRNAs/metabolism , Plant Diseases/microbiology , Plants/genetics , RNA, Plant , Verticillium/metabolismABSTRACT
Caspase-1 is an integral regulator of innate immunity, which plays a key role in inflammasome activation and the release of pro-inflammatory cytokines. The development of novel non-peptidic small molecule caspase-1 inhibitors is an important strategy for antagonizing excessively activated caspase-1 induced by inflammatory diseases, including gouty arthritis. In the present study, we identified 63 caspase-1 inhibitors, with different structures and potencies, from bioactive compound libraries. Among them, NSC697923 potently inhibited the enzymatic activity of caspase-1, with an IC50 value of 1.737 µM. This compound adopted a favorable conformation in the active pocket of caspase-1. Furthermore, NSC697923 potently decreased mature interleukin (IL)-1ß secretion in macrophages stimulated by lipopolysaccharide plus nigericin, ATP, and monosodium urate crystal. NSC697923 also inhibited NLRP3 protein expression by suppressing the NF-κB signaling pathway and the interaction between receptor interacting protein-2 (RIP2) and pro-caspase-1, thereby blocking the priming of the NLRP3 inflammasome. In addition, NSC697923 significantly inhibited caspase-1 mediated gasdermin D cleavage and pyroptosis in macrophages. In an animal model of gouty arthritis, NSC697923 effectively inhibited joint swelling, IL-1ß release, and NLRP3 inflammasome activation. Our results indicate that NSC697923 can effectively suppress NLRP3 inflammasome activation by inhibiting caspase-1, thus warranting further investigation as a potential therapeutic for treating NLRP3 inflammasome-related diseases.
Subject(s)
Arthritis, Gouty , Gout , Animals , Arthritis, Gouty/chemically induced , Arthritis, Gouty/drug therapy , Arthritis, Gouty/metabolism , Caspase 1/metabolism , Inflammasomes/metabolism , Inflammation/drug therapy , Interleukin-1beta/metabolism , NLR Family, Pyrin Domain-Containing 3 Protein/metabolism , PyroptosisABSTRACT
Kirsten rat sarcoma viral oncogene homolog (KRAS) mutations account for 35% of the genetic alterations in non-small cell lung cancer (NSCLC). The Src-homology region 2-containing protein tyrosine phosphatase 2 (SHP2), encoded by PTPN11, is closely involved in RAS downstream pathways and development of many tumors by affecting cell proliferation, differentiation, and immunity. Targeting SHP2 with small molecules may be a promising avenue for the treatment of KRAS-mutant (mut) NSCLC. Herein, hexachlorophene (HCP) was identified as a SHP2 inhibitor with an IC50 value of 5.63 ± 0.75 µM through screening of the FDA-approved drug library. HCP specifically inhibited SHP2 rather than other phosphatases. Molecular docking showed that HCP displayed an orientation favorable for nucleophilic attack in the catalytic domain of SHP2. HCP suppressed viability of multiple KRAS-mut and KRAS-wild type cells and induced senescence and apoptosis in KRAS-mut cells. Moreover, HCP reversed epithelial-mesenchymal transition to suppress metastasis in KRAS-mut cells, and inhibited the RAS/MEK/ERK and PI3K/AKT signaling pathways by suppression of SHP2 phosphorylation and formation SHP2/Grb2/Gab1/SOS1 complex. In summary, HCP can act as a specific SHP2 inhibitor to inhibit KRAS-mut NSCLC cell proliferation and metastasis and induce senescence through suppression of the RAF/MEK/ERK and PI3K/AKT pathways. HCP warrants further investigation as a new compound skeleton for the development of selective SHP2 inhibitors for the treatment of NSCLC.
Subject(s)
Carcinoma, Non-Small-Cell Lung , Lung Neoplasms , Carcinoma, Non-Small-Cell Lung/drug therapy , Carcinoma, Non-Small-Cell Lung/genetics , Carcinoma, Non-Small-Cell Lung/pathology , Cell Line, Tumor , Cell Proliferation , Hexachlorophene , Humans , Lung Neoplasms/drug therapy , Lung Neoplasms/genetics , Lung Neoplasms/pathology , Mitogen-Activated Protein Kinase Kinases/metabolism , Molecular Docking Simulation , Mutation , Phosphatidylinositol 3-Kinases/metabolism , Proto-Oncogene Proteins c-akt/metabolism , Proto-Oncogene Proteins p21(ras)/genetics , Proto-Oncogene Proteins p21(ras)/metabolism , Signal TransductionABSTRACT
Mechanical stimulation induces bone growth and remodeling by the secondary messenger, cyclic guanosine 3', 5'-monophosphate (cGMP), in osteoblasts. However, the role of cGMP in the regulation of estrogen biosynthesis, whose deficiency is a major cause of osteoporosis, remains unclear. Here, we found that the prenylated flavonoids, 3-O-methoxymethyl-7-O-benzylicaritin (13), 7-O-benzylicaritin (14), and 4'-O-methyl-8-isopentylkaempferol (15), which were synthesized using icariin analogs, promoted estrogen biosynthesis in osteoblastic UMR106 cells, with calculated EC50 values of 1.53, 3.45, and 10.57 µM, respectively. 14 and 15 increased the expression level of the bone specific promoter I.4-driven aromatase, the only enzyme that catalyzes estrogen formation by using androgens as substrates, in osteoblastic cells. 14 inhibited phosphodiesterase 5 (PDE5), stimulated intracellular cGMP level and promoted osteoblast cell differentiation. Inhibition of cGMP dependent-protein kinase G (PKG) abolished the stimulatory effect of 14 on estrogen biosynthesis and osteoblast cell differentiation. Further, PKG activation by 14 stimulated the activity of SHP2 (Src homology 2 domain-containing tyrosine phosphatase 2), thereby activating Src and ERK (extracellular signal-regulated kinase) signaling and increasing ERK-dependent aromatase expression in osteoblasts. Our findings reveal a previously unknown role of cGMP in the regulation of estrogen biosynthesis in the bone. These results support the further development of 14 as a PKG-activating drug to mimic the anabolic effects of mechanical stimulation of bone in the treatment of osteoporosis.
Subject(s)
Aromatase/metabolism , Cyclic Nucleotide Phosphodiesterases, Type 5/metabolism , Estrogens/metabolism , Osteoblasts/metabolism , 3T3 Cells , Animals , Cell Differentiation , Cell Line , Cell Survival , Cyclic AMP/metabolism , Cyclic GMP/metabolism , Cyclic GMP-Dependent Protein Kinases/metabolism , Flavonoids/chemistry , HEK293 Cells , Humans , Mice , Molecular Docking Simulation , Osteoblasts/cytology , Osteoporosis , Protein Tyrosine Phosphatase, Non-Receptor Type 11/metabolism , Stress, MechanicalABSTRACT
Flowering plants and mammals contain imprinted genes that are primarily expressed in the endosperm and placenta in a parent-of-origin manner. In this study, we show that early activation of the geminivirus genes C2 and C3 in Arabidopsis (Arabidopsis thaliana) plants, encoding a viral suppressor of RNA interference and a replication enhancer protein, respectively, is correlated with the transient vegetative expression of VARIANT IN METHYLATION5 (VIM5), an endosperm imprinted gene that is conserved in diverse plant species. VIM5 is a ubiquitin E3 ligase that directly targets the DNA methyltransferases MET1 and CMT3 for degradation by the ubiquitin-26S proteasome proteolytic pathway. Infection with Beet severe curly top virus induced VIM5 expression in rosette leaf tissues, possibly via the expression of the viral replication initiator protein, leading to the early activation of C2 and C3 coupled with reduced symmetric methylation in the C2-3 promoter and the onset of disease symptoms. These findings demonstrate how this small DNA virus recruits a host imprinted gene for the epigenetic activation of viral gene transcription. Our findings reveal a distinct strategy used by plant pathogens to exploit the host machinery in order to inhibit methylation-mediated defense responses when establishing infection.
Subject(s)
Arabidopsis/genetics , Arabidopsis/virology , Geminiviridae/pathogenicity , Plant Diseases/virology , Arabidopsis Proteins/genetics , Arabidopsis Proteins/metabolism , DNA (Cytosine-5-)-Methyltransferases/genetics , DNA (Cytosine-5-)-Methyltransferases/metabolism , DNA Methylation , DNA-Cytosine Methylases/genetics , DNA-Cytosine Methylases/metabolism , Epigenesis, Genetic , Gene Expression Regulation, Plant , Genomic Imprinting , Host-Pathogen Interactions/genetics , Plant Diseases/genetics , Plant Leaves/genetics , Plant Leaves/virology , Plants, Genetically Modified , Promoter Regions, Genetic , Ubiquitin-Protein Ligases/genetics , Ubiquitin-Protein Ligases/metabolism , Viral Proteins/geneticsABSTRACT
Male and female gametophytes are generated from micro- or megaspore mother cells through consecutive meiotic and mitotic cell divisions. Defects in these divisions often result in gametophytic lethality. Gametophytic lethality was also reported when genes encoding ribosome-related proteins were mutated. Although numerous ribosomal proteins (RPs) have been identified in plants based on homology with their yeast and metazoan counterparts, how RPs are regulated, e.g., through dynamic subcellular targeting, is unknown. We report here that an Arabidopsis (Arabidopsis thaliana) importin ß, KETCH1 (karyopherin enabling the transport of the cytoplasmic HYL1), is critical for gametogenesis. Karyopherins are molecular chaperones mediating nucleocytoplasmic protein transport. However, the role of KETCH1 during gametogenesis is independent of HYPONASTIC LEAVES 1 (HYL1), a previously reported KETCH1 cargo. Instead, KETCH1 interacts with several RPs and is critical for the nuclear accumulation of RPL27a, whose mutations caused similar gametophytic defects. We further showed that knocking down KETCH1 caused reduced ribosome biogenesis and translational capacity, which may trigger the arrest of mitotic cell cycle progression and lead to gametophytic lethality.
Subject(s)
Arabidopsis Proteins/metabolism , Arabidopsis/growth & development , Arabidopsis/metabolism , Cell Nucleus/metabolism , Gametogenesis, Plant , Karyopherins/metabolism , Ribosomal Proteins/metabolism , Arabidopsis/genetics , Arabidopsis/ultrastructure , Cell Cycle Checkpoints , Cell Nucleus/ultrastructure , Down-Regulation , Gene Expression Regulation, Plant , Loss of Function Mutation/genetics , Ovule/metabolism , Ovule/ultrastructure , Pollen/growth & development , Pollen/ultrastructure , Protein Binding , Protein Biosynthesis , RNA-Binding Proteins/metabolism , Ribosomes/metabolism , Seeds/metabolism , Seeds/ultrastructureABSTRACT
BACKGROUND: Multiple organ dysfunction syndromes (MODS) is reported as a leading cause of mortality in intensive care units. Recently, continuous blood purification (CBP) has been mostly applied for MODS treatment. Thus, the purpose of this study was to investigate the effects of CBP on plasma phospholipid level in patients with MODS. METHODS: A total of 126 patients with MODS and 120 healthy people were collected. The serum cytokine levels, blood biochemical parameters, and blood gas indexes were detected, and the correlation among phospholipid compounds with serum cytokine levels, blood biochemical parameters, and blood gas indexes was analyzed. RESULTS: Before CBP, levels of body temperature, RR, HR, CVP, IL-6, IL-10, TNF-α, BUN, SCr, PaCO2 , SM747, and LPC540 were obviously higher, and pH, HCO3- , PaO2 , SaO2 , PE750, PI885, PC792, PC826, PC830, PC854, PC802, and PG747 were lower in the MODS group than those in the control group. During CBP, the MODS group had gradually declined RR, CVP, levels of IL-6, IL-10 and TNF-α, BUN, SCr, PaCO2 , SM747, and LPC540 and increased HCO3- , PaO2 and SaO2 , PE750, PI885, PC792, PC826, PC830, PC854, PC802, and PG747. Besides, levels of PE750, PI885, PC792, PC826, PC830, PC854, PC802, and PG747 had an obvious negative correlation with levels of TNF-α, IL-10, IL-6, BUN, SCr, and PaCO2 , and a significant positive correlation with levels of HCO3- , PaO2 , and SaO2 . CONCLUSION: CBP could effectively ameliorate clinical signs of patients with MODS and correct the plasma phospholipid levels.
ABSTRACT
Mitochondrial genome plays a central role in aging, cancer, apoptosis and metabolism. We sequenced a complete mitochondrial genome sequence of a rat chronic obstructive pulmonary disease rat strain for the first time. The total length of the mitochondrial genome was 16,310 bp and coding 13 protein-coding genes, two ribosomal RNA genes, 22 transfer RNA genes. This genome describing information will supply the potential use of mtDNA mutations as markers in cancer.
Subject(s)
Genome, Mitochondrial , Pulmonary Disease, Chronic Obstructive/genetics , Animals , Codon, Initiator , Codon, Terminator , DNA, Mitochondrial/chemistry , DNA, Mitochondrial/isolation & purification , DNA, Mitochondrial/metabolism , Disease Models, Animal , Electron Transport Complex IV/chemistry , Electron Transport Complex IV/genetics , Electron Transport Complex IV/metabolism , NADH Dehydrogenase/chemistry , NADH Dehydrogenase/genetics , NADH Dehydrogenase/metabolism , Open Reading Frames/genetics , Pulmonary Disease, Chronic Obstructive/pathology , RNA, Ribosomal/chemistry , RNA, Ribosomal/genetics , RNA, Transfer/chemistry , RNA, Transfer/genetics , Rats , Sequence Analysis, DNAABSTRACT
The study describes the development of polylactide-tocopheryl polyethylene glycol 1000 succinate (PLA-TPGS)-based nanosystem as a carrier of crizotinib (CZT) to achieve superior anticancer efficacy in lung cancer therapy. We have demonstrated that block copolymer and hydrophobic drug is capable of self-assembling into a very stable nanocarrier, with suitable properties that allow their application for cancer drug delivery. Drug release study showed a sustained release pattern as a result of entrapment in the hydrophobic core of micelles. CZT/PT NP showed a noticeable cytotoxic effect in NCIH3122 lung cancer cells in a dose-dependent manner. Furthermore, morphological imaging and Live/Dead assay revealed a superior anticancer efficacy for nanoformulations. The polymeric nanoparticle showed a predominant presence in the cytoplasmic region of cell, indicating a typical endocytosis-mediated cellular uptake. The annexin V/PI staining-based apoptosis assay showed a remarkable ~40 % apoptosis (early and late apoptosis cells) comparing to only ~25 % apoptosis by free CZT. Taken together, Vitamin E TPGS-modified PLA nanoparticles would be a potential drug delivery system to increase the chemotherapeutic efficacy of CZT in lung cancer chemotherapy.
Subject(s)
Lung Neoplasms/drug therapy , Nanoparticles/administration & dosage , Polymers/administration & dosage , Pyrazoles/administration & dosage , Pyridines/administration & dosage , Antineoplastic Agents/administration & dosage , Apoptosis/drug effects , Cell Line, Tumor , Crizotinib , Drug Carriers/administration & dosage , Drug Delivery Systems/methods , Humans , Polyethylene Glycols/administration & dosage , Succinates/administration & dosageABSTRACT
Cytosine methylation is one of epigenetic information marked on the DNA sequence. In plants, small interfering RNAs (siRNAs) target homologous genomic DNA sequences for cytosine methylation. This process, known as RNA-directed DNA methylation (RdDM), plays an important role in transposon control, regulation of gene expression and virus resistance. In this paper, we demonstrate that the C2 protein encoded by a geminivirus (beet severe curly top virus, BSCTV) mediated a decrease in DNA methylation of repeat regions in the promoters of ACD6, an upstream regulator of the salicylic acid defense pathway, and GSTF14, an endogenous gene of the glutathione S-transferase superfamily that is implicated in numerous stress responses. C2-mediated decreases in DNA methylation reduced accumulation of the siRNAs derived from the promoter repeats and enhanced the steady-state expression of both ACD6 and GSTF14 transcripts. Reduced accumulation of BSCTV-derived siRNAs was detected in BSCTV-infected plants, but not in plants infected with C2-deficient BSCTV (c2(- ) BSCTV). C2 protein exhibited no siRNA-binding activity. Instead, our results revealed that C2 protein-mediated decreases in DNA methylation appeared to affect the production of siRNAs that are required for targeting and reinforcing RdDM, a process that activated expression of defense-related genes that are normally dampened by these siRNAs in the host plants. However, C2-dependent reduction in virus-derived siRNAs also benefits the viruses by disrupting the feedback loop reinforcing DNA methylation-mediated antiviral silencing.
Subject(s)
Arabidopsis/genetics , Arabidopsis/virology , DNA Methylation , Geminiviridae/pathogenicity , Gene Expression Regulation, Plant , Host-Pathogen Interactions/genetics , Viral Proteins/metabolism , Ankyrins/genetics , Arabidopsis Proteins/genetics , Glutathione Transferase/genetics , Glutathione Transferase/metabolism , Plants, Genetically Modified , Promoter Regions, Genetic , RNA, Small Interfering/genetics , Salicylic Acid/metabolism , Viral Proteins/geneticsABSTRACT
Taking the dominant tree species Quercus mongolica in natural coniferous-broadleaved mixed forest in Changbai Mountains as test object, this paper studied the variations of leaf dry mass per unit area (LMA), leaf carbon (C), nitrogen (N), and phosphorus (P) contents per unit mass and per unit area, as well as the leaf N and P resorption efficiency and use efficiency at upper and lower canopy positions during growth season (from June to October). In the growth season, and at both upper and lower canopy positions, the LMA and leaf C content per unit area had obvious monthly fluctuation, the leaf N and P contents per unit area had the similar monthly variation trend with the leaf N and P contents per unit mass, but the leaf N and P resorption efficiency per unit mass had no significant difference with the leaf N and P resorption efficiency per unit area. The leaf N resorption efficiency and use efficiency were less affected by canopy position, but the leaf P resorption efficiency and use efficiency were higher at upper canopy than at lower canopy. Under the scenario of future climate change, the higher survival and competitive capabilities of Q. mongolica would benefit the nutrient cycling in the test forest ecosystem.
Subject(s)
Nitrogen/metabolism , Plant Leaves/metabolism , Quercus/metabolism , Absorption , Carbon/metabolism , Ecosystem , Phosphorus/metabolism , Quercus/growth & developmentABSTRACT
Midkine is a heparin-binding growth factor, which plays important roles in the regulation of cell growth and differentiation. The non-tagged recombinant human midkine (rhMK) is therefore required to facilitate its functional studies of this important growth factor. In the present work, rhMK was expressed in Escherichia coli (E. coli) BL21 (DE3). The expression of midkine was efficiently induced by isopropyl-beta-D-thiogalactopyranoside (IPTG). After sonication, midkine was recovered in an insoluble form, and was dissolved in guanidine hydrochloride buffer. Renaturation of the denatured protein was carried out in the defined protein refolding buffer, and the refolded protein was purified using S-Sepharose ion-exchange chromatography. The final preparation of the rhMK was greater than 98% pure as measured by sodium dodecylsulfate-polyacrylamid gel electrophoresis (SDS-PAGE) and reverse phase high performance liquid chromatography (RP-HPLC). The purified rhMK enhanced the proliferation of NIH3T3 cells.
Subject(s)
Cytokines/biosynthesis , Recombinant Proteins/biosynthesis , Animals , Base Sequence , Cell Proliferation/drug effects , Cytokines/genetics , Cytokines/isolation & purification , Cytokines/pharmacology , Escherichia coli/genetics , Humans , Mice , Midkine , Molecular Sequence Data , NIH 3T3 Cells , Recombinant Proteins/isolation & purificationABSTRACT
It has been reported that plant virus-derived small interfering RNAs (vsiRNAs) originated predominantly from structured single-stranded viral RNA of a positive single-stranded RNA virus replicating in the cytoplasm and from the nuclear stem-loop 35S leader RNA of a double-stranded DNA (dsDNA) virus. Increasing lines of evidence have also shown that hierarchical actions of plant Dicer-like (DCL) proteins are required in the biogenesis process of small RNAs, and DCL4 is the primary producer of vsiRNAs. However, the structures of such single-stranded viral RNA that can be recognized by DCLs remain unknown. In an attempt to determine these structures, we have cloned siRNAs derived from the satellite RNA (satRNA) of Cucumber mosaic virus (CMV-satRNA) and studied the relationship between satRNA-derived siRNAs (satsiRNAs) and satRNA secondary structure. satsiRNAs were confirmed to be derived from single-stranded satRNA and are primarily 21 (64.7%) or 22 (22%) nucleotides (nt) in length. The most frequently cloned positive-strand satsiRNAs were found to derive from novel hairpins that differ from the structure of known DCL substrates, miRNA and siRNA precursors, which are prevalent stem-loop-shaped or dsRNAs. DCL4 was shown to be the primary producer of satsiRNAs. In the absence of DCL4, only 22-nt satsiRNAs were detected. Our results suggest that DCL4 is capable of accessing flexibly structured single-stranded RNA substrates (preferably T-shaped hairpins) to produce satsiRNAs. This result reveals that viral RNA of diverse structures may stimulate antiviral DCL activities in plant cells.
Subject(s)
Arabidopsis/immunology , Cucumber Mosaic Virus Satellite/genetics , Cucumber Mosaic Virus Satellite/metabolism , Cucumovirus/immunology , Plant Diseases/immunology , RNA, Small Interfering/metabolism , Ribonucleases/metabolism , Arabidopsis/virology , Base Sequence , Binding Sites , Cucumber Mosaic Virus Satellite/chemistry , Cucumovirus/genetics , Models, Molecular , Molecular Sequence Data , Nucleic Acid Conformation , Plant Diseases/virology , RNA Interference , RNA, Small Interfering/genetics , Ribonuclease III , Ribonucleases/immunologyABSTRACT
A bacterium capable of utilizing fenpropathrin as sole carbon source was isolated from activated sludge collected from wastewater treating system of a pesticide manufacturer. This bacterium was identified as Sphingomonas sp. according to its physiological & biochemical analysis and the similarity analysis of its 16S rDNA sequence (GenBank Accession No. DQ177525). This bacterium could degrade 99.8% of 20 mg/L fenpropathrin in 24h. The optimal pH and temperature for the degradation were 7.0 and 30 degrees C, respectively. The degradation speed was related positively to initial inoculum size. The enzyme distribution experiment showed that the degrading-enzyme in the bacterium was endoenzyme.
Subject(s)
Pyrethrins/metabolism , Sphingomonas/isolation & purification , Sphingomonas/metabolism , Biodegradation, Environmental , Hydrogen-Ion Concentration , Insecticides/metabolism , Microscopy, Electron , Molecular Sequence Data , Phylogeny , RNA, Ribosomal, 16S/genetics , Sequence Analysis, DNA , Sphingomonas/genetics , TemperatureABSTRACT
A facilitative and efficient promoter-trapping vector, pUC-mpd, was constructed with the promoterless methyl parathion hydrolase gene as the reporter. This reporter gene is easily used to clone promoters with different promoting strength on selective plates. Promoter regions of the ytkA and ywoF genes with strong promoting and signal peptide functions were cloned from the Bacillus subtilis 168 genomic promoter library with this vector.