RESUMEN
Excessive oxidative stress impairs cartilage matrix metabolism balance, significantly contributing to osteoarthritis (OA) development. Celastrol (CSL), a drug derived from Tripterygium wilfordii, has recognized applications in the treatment of cancer and immune system disorders, yet its antioxidative stress mechanisms in OA remain underexplored. This study aimed to substantiate CSL's chondroprotective effects and unravel its underlying mechanisms. We investigated CSL's impact on chondrocytes under both normal and inflammatory conditions. In vitro, CSL mitigated interleukin (IL)-1ß-induced activation of proteinases and promoted cartilage extracellular matrix (ECM) synthesis. In vivo, intra-articular injection of CSL ameliorated cartilage degeneration and mitigated subchondral bone lesions in OA mice. Mechanistically, it was found that inhibiting nuclear factor erythroid 2-related factor 2 (NRF2) abrogated CSL-mediated antioxidative functions and exacerbated the progression of OA. This study is the first to elucidate the role of CSL in the treatment of OA through the activation of NRF2, offering a novel therapeutic avenue for arthritis therapy.
Asunto(s)
Factor 2 Relacionado con NF-E2 , Osteoartritis , Ratones , Animales , Factor 2 Relacionado con NF-E2/genética , Factor 2 Relacionado con NF-E2/metabolismo , Antioxidantes/farmacología , Antioxidantes/metabolismo , Osteoartritis/tratamiento farmacológico , Osteoartritis/metabolismo , Osteoartritis/patología , Triterpenos Pentacíclicos/farmacología , Triterpenos Pentacíclicos/metabolismo , Condrocitos , Interleucina-1betaRESUMEN
The unicellular freshwater green alga Haematococcus pluvialis has attracted much research attention due to its biosynthetic ability for large amounts of astaxanthin, a blood-red ketocarotenoid that is used in cosmetics, nutraceuticals, and pharmaceuticals. Recently, numerous studies have investigated the functions of natural astaxanthin; however, the high cost of the production of astaxanthin from H. pluvialis cultures restricts its commercial viability. There is an urgent need to fulfill commercial demands by increasing astaxanthin accumulation from H. pluvialis cultures. In this study, we discovered that treatment of H. pluvialis cultures at the beginning of the macrozooid stage (day 0) with 1 µM rac-GR24, a synthetic analogue of strigolactones (a class of phytohormones), led to significant increases in biomass [up to a maximum dry cell weight (DCW) of 0.53 g/L] during the macrozooid stage and astaxanthin (from 0.63 to 5.32% of DCW) during the hematocyst stage. We elucidated that this enhancement of biomass accumulation during the macrozooid stage by rac-GR24 is due to its increasing CO2 utilization efficiency in photosynthesis and carbohydrate biosynthesis. We also found that rac-GR24 stimulated the overproduction of nicotinamide adenine dinucleotide phosphate (NADPH) and antioxidant enzymes in H. pluvialis cultures, which alleviated the oxidative damage caused by reactive oxygen species generated during the hematocyst stage due to the exhaustion of nitrogen supplies. Moreover, rac-GR24 treatment of H. pluvialis synergistically altered the activity of the pathways of fatty acid biosynthesis and astaxanthin esterification, which resulted in larger amounts of astaxanthin being generated by rac-GR24-treated cultures than by controls. In summary, we have developed a feasible and economic rac-GR24-assisted strategy that increases the amounts of biomass and astaxanthin generated by H. pluvialis cultures, and have provided novel insights into the mechanistic roles of rac-GR24 to achieve these effects.
Asunto(s)
Chlorophyceae , Biomasa , Suplementos Dietéticos , XantófilasRESUMEN
Phosphorus (P) limitation affects phytoplankton growth and population size in aquatic systems, and consequently limits aquatic primary productivity. Plants have evolved a range of metabolic responses to cope with P limitation, such as accumulation of purple acid phosphatases (PAPs) to enhance acquisition of phosphates. However, it remains unknown whether algae have evolved a similar mechanism. In this study, we examined the role of PAPs in the model microalga Phaeodactylum tricornutum. Expression of PAP1 was enhanced in P. tricornutum cells grown on organophosphorus compared to inorganic phosphate. PAP1 overexpression improved cellular growth and biochemical composition in a growth-phase dependent manner. PAP1 promoted growth and photosynthesis during growth phases and reallocated carbon flux towards lipogenesis during the stationary phase. PAP1 was found to be localized in the endoplasmic reticulum and it orchestrated the expression of genes involved in key metabolic pathways and translocation of inorganic P (Pi), thereby improving energy use, reducing equivalents and antioxidant potential. RNAi of PAP1 induced expression of its homolog PAP2, thereby compensating for the Pi scavenging activity of PAP1. Our results demonstrate that PAP1 brings about sequential regulation of metabolism, and provide novel insights into algal phosphorus metabolism and aquatic primary productivity.
Asunto(s)
Diatomeas , Fosfatasa Ácida/metabolismo , Diatomeas/genética , Diatomeas/metabolismo , Hidrólisis , Fósforo , FotosíntesisRESUMEN
Aims: The study aimed to understand the role and the core values of pharmacists and the professional expectations of medical staff for pharmacists in treating COVID-19 patients from the perspectives of the frontline medical staff. The findings help to understand and provide a reference for the career growth path of future pharmacists. Methods: A phenomenological method was used to conduct in-depth interviews with frontline medical staff working in isolation wards during COVID-19. The interview data were analyzed, and the themes were extracted. Results: Pharmacists played a positive role in ensuring the supply of non-routinely stocked drugs, including traditional Chinese medicine preventative preparations, providing drug information and medication consultation for complex patients, and identifying adverse drug reactions. However, at present, the integration of pharmacists and nurses is poor with inadequate communication, and the pharmaceutical care activities provided to physicians were still not comprehensive. Conclusions: The level of pharmaceutical care provided by pharmacists needs to be further strengthened. Frontline medical teams generally have high professional expectations for pharmacists, including expecting pharmacists to become drug therapy experts. They expect pharmacists to fully participate in clinical decision-making, especially playing a central role in managing drug interactions, contraindications, and other clinical uses of drugs.
Asunto(s)
COVID-19 , Farmacéuticos , Humanos , Cuerpo Médico , Motivación , SARS-CoV-2RESUMEN
Tannase (E.C. 3.1.1.20) is hypothesized to be involved in the metabolism of gallates and gallic acid (GA) in pu-erh tea fermentation. In this work, we measured tannase in Aspergillus niger fermented tea leaves and confirmed the production of fungal tannase during pu-erh tea fermentation. A decrease in catechin and theaflavin gallates and a significant increase in GA content and the relative peak areas of ethyl gallate, procyanidin A2, procyanidin B2, procyanidin B3, catechin-catechin-catechin, epiafzelechin, and epicatechin-epiafzelechin [variable importance in the projection (VIP) > 1.0, p < 0.05, and fold change (FC) > 1.5] were observed using high performance liquid chromatography (HPLC) and metabolomics analysis of tea leaves fermented or hydrolyzed by tannase. In vitro assays showed that hydrolysis by tannase or polymerization of catechins increased the antioxidant activity of tea leaves. In summary, we identified a metabolic pathway for gallates and their derivatives in tea leaves hydrolyzed by tannase as well as associated changes in gallate and GA concentrations caused by fungal tannase during pu-erh tea fermentation.
Asunto(s)
Aspergillus niger/metabolismo , Camellia sinensis/microbiología , Hidrolasas de Éster Carboxílico/metabolismo , Proteínas Fúngicas/metabolismo , Ácido Gálico/metabolismo , Aspergillus niger/química , Aspergillus niger/enzimología , Camellia sinensis/química , Camellia sinensis/metabolismo , Hidrolasas de Éster Carboxílico/química , Cromatografía Líquida de Alta Presión/métodos , Fermentación , Proteínas Fúngicas/química , Ácido Gálico/química , Metabolómica/métodos , Hojas de la Planta/química , Hojas de la Planta/metabolismo , Hojas de la Planta/microbiologíaRESUMEN
The effects of five natural products from Chinese herbs including evodiamine, curcumin, 4-methoxysalicylaldehyde, esculin hydrate, and gramine on the growth of Chattonella marina, one of the most noxious red tide algae, were observed. Among them, gramine exhibited the highest inhibitory rate with LC50, 96h of 0.51 mg/l. After exposure to gramine, the activities of superoxide dismutase (SOD) and catalase (CAT), and content of malondialdehyde (MDA) increased in C. marina, suggesting that gramine could induce microalgae oxidative stress. In addition, chlorophyll a and the maximum quantum yield of photosynthesis (Fv/Fm) decreased following exposure to gramine, indicating the inhibition of photosynthesis activity in the microalgae. Combined with the fast inhibition against the algal cells and environmentally friendly character of gramine, we proposed that gramine might be a potential algaecide against marine harmful algae and that the oxidative damage and photosynthesis inhibition might be responsible for the toxicity of gramine on harmful algae.
Asunto(s)
Productos Biológicos/farmacología , Medicamentos Herbarios Chinos/farmacología , Estramenopilos/efectos de los fármacos , Alcaloides/farmacología , Benzaldehídos/farmacología , Catalasa/metabolismo , Clorofila/metabolismo , Curcumina/farmacología , Floraciones de Algas Nocivas/efectos de los fármacos , Alcaloides Indólicos , Malondialdehído/metabolismo , Microalgas/metabolismo , Oxidación-Reducción , Estrés Oxidativo , Fotosíntesis/efectos de los fármacos , Quinazolinas/farmacología , Estramenopilos/fisiología , Superóxido Dismutasa/metabolismoRESUMEN
Phosphorus (P) is an essential macronutrient for the survival of marine phytoplankton. In the present study, phytoplankton response to phosphorus limitation was studied by proteomic profiling in diatom Phaeodactylum tricornutum in both cellular and molecular levels. A total of 42 non-redundant proteins were identified, among which 8 proteins were found to be upregulated and 34 proteins were downregulated. The results also showed that the proteins associated with inorganic phosphate uptake were downregulated, whereas the proteins involved in organic phosphorus uptake such as alkaline phosphatase were upregulated. The proteins involved in metabolic responses such as protein degradation, lipid accumulation and photorespiration were upregulated whereas energy metabolism, photosynthesis, amino acid and nucleic acid metabolism tend to be downregulated. Overall our results showed the changes in protein levels of P. tricornutum during phosphorus stress. This study preludes for understanding the role of phosphorous in marine biogeochemical cycles and phytoplankton response to phosphorous scarcity in ocean. It also provides insight into the succession of phytoplankton community, providing scientific basis for elucidating the mechanism of algal blooms.
Asunto(s)
Organismos Acuáticos/genética , Diatomeas/genética , Fósforo/metabolismo , Proteómica , Organismos Acuáticos/metabolismo , Diatomeas/metabolismo , Perfilación de la Expresión Génica , Nitrógeno , Biosíntesis de Proteínas/genética , Estrés Fisiológico/genéticaRESUMEN
Phosphorus is an important macronutrient. To understand the molecular and cellular responses to phosphorus stress better, transcriptome profiling in combination with biochemical investigations was conducted in the model diatom Phaeodactylum tricornutum. Out of 10 402 predicted genes, 2491 and 405 genes were significantly upregulated or downregulated respectively. Unsurprisingly, genes associated with phosphate uptake were upregulated, such as the phosphate transporters and alkaline phosphatases. Genes encoding stress-shock proteins were accordingly upregulated, including genes associated with stress-responsive proteins, signal transduction and secondary metabolism. Additionally, genes related to protein translation, carbon fixation, glycolysis and the citric acid cycle were also upregulated. Genes associated with gene transcription were downregulated, thereby resulting in the upregulation of translation to compensate for the limited supply of messenger RNA. The downregulation of genes related to ß-oxidation could contribute to the accumulation of fatty acids. Accordingly, triacylglycerols, which are important for energy storage, were determined to increase by 1.65-fold. Intracellular membranes, other than chloroplast membranes, tended to be dispersed; this finding was in accordance with the increased transcription of a total of 11 genes encoding putative phospholipases. Taken together, this work revealed the coordination of multiple metabolic pathways and certain key genes in the adaptation of P. tricornutum to phosphorus stress.
Asunto(s)
Diatomeas/metabolismo , Fósforo/metabolismo , Estrés Fisiológico , Adaptación Fisiológica , Ciclo del Carbono , Ciclo del Ácido Cítrico , Diatomeas/genética , Perfilación de la Expresión Génica , Glucólisis , Membranas Intracelulares/metabolismo , Metabolismo de los Lípidos , Proteínas de Transporte de Fosfato/genética , Proteínas de Transporte de Fosfato/metabolismo , ARN Mensajero/genética , ARN Mensajero/metabolismo , Análisis de Secuencia de ARN , Transcriptoma , Regulación hacia ArribaRESUMEN
BACKGROUND: The production of watermelon in China has been seriously hampered by fruit blotch disease and limited control measures are now applied. Chitosan has been employed to control a variety of plant diseases and is considered to be the most promising biochemical to control this disease. RESULTS: The in vitro antibacterial effect of chitosan and its ability in protection of watermelon seedlings from bacterial fruit blotch were evaluated. Results showed that three types of chitosan, in particular, chitosan A at 0.40 mg mL⻹ significantly inhibited the growth of Acidovorax citrulli. The antibacterial activity of chitosan A was affected by chitosan concentration and incubation time. The direct antibacterial activity of chitosan may be attributed to membrane lysis evidenced by transmission electron microscopic observation. The disease index of watermelon seedlings planted in soil and the death rate of seedlings planted in perlite were significantly reduced by chitosan A at 0.40 mg mL⻹ compared to the pathogen control. Fresh and dry weight of watermelon seedlings planted in soil was increased by chitosan seed treatment, but not by chitosan leaf spraying. CONCLUSION: The results indicated that chitosan solution may have a potential in controlling bacterial fruit blotch of watermelon.
Asunto(s)
Agroquímicos/farmacología , Antibacterianos/farmacología , Quitosano/farmacología , Citrullus/efectos de los fármacos , Comamonadaceae/efectos de los fármacos , Enfermedades de las Plantas/prevención & control , Plantones/efectos de los fármacos , Acetilación , Agroquímicos/química , Óxido de Aluminio , Antibacterianos/química , Membrana Celular/efectos de los fármacos , Membrana Celular/ultraestructura , China , Quitosano/química , Citrullus/enzimología , Citrullus/crecimiento & desarrollo , Citrullus/microbiología , Recuento de Colonia Microbiana , Comamonadaceae/crecimiento & desarrollo , Comamonadaceae/aislamiento & purificación , Comamonadaceae/ultraestructura , Frutas/efectos de los fármacos , Frutas/enzimología , Frutas/crecimiento & desarrollo , Frutas/microbiología , Pruebas de Sensibilidad Microbiana , Viabilidad Microbiana , Peso Molecular , Concentración Osmolar , Enfermedades de las Plantas/microbiología , Proteínas de Plantas/metabolismo , Plantones/enzimología , Plantones/crecimiento & desarrollo , Plantones/microbiología , Semillas/efectos de los fármacos , Semillas/enzimología , Semillas/crecimiento & desarrollo , Semillas/microbiología , Dióxido de Silicio , SueloRESUMEN
Pectins are complex polysaccharides that are essential components of the plant cell wall. In this study, a novel putative Arabidopsis S-adenosyl-L-methionine (SAM)-dependent methyltransferase, termed QUASIMODO 3 (QUA3, At4g00740), has been characterized and it was demonstrated that it is a Golgi-localized, type II integral membrane protein that functions in methylesterification of the pectin homogalacturonan (HG). Although transgenic Arabidopsis seedlings with overexpression, or knock-down, of QUA3 do not show altered phenotypes or changes in pectin methylation, this enzyme is highly expressed and abundant in Arabidopsis suspension-cultured cells. In contrast, in cells subjected to QUA3 RNA interference (RNAi) knock-down there is less pectin methylation as well as altered composition and assembly of cell wall polysaccharides. Taken together, these observations point to a Golgi-localized QUA3 playing an essential role in controlling pectin methylation and cell wall biosynthesis in Arabidopsis suspension cell cultures.
Asunto(s)
Proteínas de Arabidopsis/metabolismo , Arabidopsis/enzimología , Pared Celular/metabolismo , Metiltransferasas/metabolismo , Secuencia de Aminoácidos , Arabidopsis/clasificación , Arabidopsis/genética , Arabidopsis/metabolismo , Proteínas de Arabidopsis/química , Proteínas de Arabidopsis/genética , Pared Celular/química , Pared Celular/enzimología , Pared Celular/genética , Células Cultivadas , Evolución Molecular , Aparato de Golgi/genética , Metilación , Metiltransferasas/química , Metiltransferasas/genética , Datos de Secuencia Molecular , Pectinas/biosíntesis , Filogenia , Transporte de Proteínas , Alineación de SecuenciaRESUMEN
OBJECTIVE: To explore the effects of some membrane lipids on the hemolysis induced by hemolytic toxin from Karenia mikimotoi. METHODS: Effects of exogenous membrane lipids such as lecithin, sphingomyelin, L-alpha-phosphatidic acid,cholesterol and gangliosides on the hemolysis induced by the hemolytic toxin were observed. The sensitivities of some erythrocytes from different animals such as rabbit, rat and fish to the hemolytic toxin were evaluated. The total gangliosides in different erythrocytes membrane were detected by colorimetry. RESULTS: Only gangliosides significantly inhibited the hemolysis of the hemolytic toxin from K. mikimotoi (P <0.05). Hemolytic percentages decreased to 16.05% after 10 min addition of ganglioside, while those of control were 35.65%. The rabbit red blood cell was the most sensitive to the hemolytic toxin. The hemolytic percentages of rabbit erythrocyte were higher than those of rat (P < 0.05) and fish (P < 0.01). The amounts of lipid-bind sialic acid (LBSA) on frozen dried membrane of rabbit were 672.08 microg/g,and were higher than those of rat (585.97 microg/g) (P < 0.05) and that of fish (431.52 microg/g) (P < 0.01). CONCLUSION: Exogenous gangliosides could have a potent inhibition on the hemolysis induced by hemolytic toxin from K. mikimotoi. There was a significant correlation between the sensitivities of different erythrocytes to the hemolytic toxin and the amount of ganglioside on different erythrocytes membrane.
Asunto(s)
Dinoflagelados/metabolismo , Eritrocitos/efectos de los fármacos , Hemólisis/efectos de los fármacos , Lípidos de la Membrana/farmacología , Toxinas Biológicas/toxicidad , Animales , Peces , Gangliósidos/farmacología , Lecitinas/farmacología , Conejos , Ratas , Esfingomielinas/farmacología , Toxinas Biológicas/biosíntesisRESUMEN
The wood sawdust from Chinese fir (Cunninghamia lanceolata) exhibited stronger inhibition on the growth of Alexandrium tamarense than those from alder (Alnus cremastogyne), pine (Pinus massoniana), birch (Betula alnoides) and sapele (Entandrophragma cylindricum). The water extract, acetone-water extract and essential oil from fir sawdust were all shown to inhibit the growth of A. tamarense. The inhibition of fir essential oil was the strongest among all the above wood sources while the half effective concentration was only 0.65 mg/L. These results suggested that the fir essential oil may play an important role in the algicidal effect of Chinese fir.
Asunto(s)
Cunninghamia/química , Dinoflagelados/efectos de los fármacos , Extractos Vegetales/farmacología , Animales , Madera/químicaRESUMEN
Cytosolic 10-kDa acyl-CoA-binding proteins (ACBPs) function in the storage and intracellular transport of acyl-CoA esters in eukaryotes. Fatty acids synthesized de novo in plant chloroplasts are exported as oleoyl-CoA and palmitoyl-CoA esters. In Arabidopsis, other than the 10-kDa ACBP, there exists five larger ACBPs (ACBP1 to ACBP5) of which homologues have not been characterized in other organisms. To investigate the significance of this gene family, we have attempted to subcellularly localize them and compare their acyl-CoA-binding affinities. We have previously shown that Arabidopsis ACBP1 and ACBP2 are membrane-associated proteins while ACBP4 and ACBP5 contain kelch motifs. Here, to localize ACBP3, we have expressed ACBP3-red fluorescent protein (DsRed2) from the CaMV 35S promoter. ACBP3-DsRed was localized extracellularly in transiently expressed tobacco BY-2 cells and onion epidermal cells. The function of the acyl-CoA-binding domain in ACBP3 was investigated by in vitro binding assays using (His)(6)-ACBP3, which was observed to bind [(14)C]arachidonyl-CoA with high affinity in comparison to [(14)C]palmitoyl-CoA and [(14)C]oleoyl-CoA. To identify the residues functional in binding, five mutants with single amino acid substitutions in the acyl-CoA-binding domain of (His)(6)-ACBP3 and (His)(6)-ACBP1 (which also binds [(14)C]arachidonyl-CoA) were generated by site-directed mutagenesis. Binding assays with arachidonyl-CoA revealed that replacement of a conserved R residue (R150A in ACBP1 and R284A in ACBP3), disrupted binding. In contrast, other substitutions in ACBP1 (Y126A, K130A, K152A and Y171A) and in ACBP3 (F260A, K264A, K286A and Y305A) did not affect arachidonyl-CoA binding, unlike their equivalents in (His)(6)-ACBP2, (His)(6)-ACBP4 and (His)(6)-ACBP5, which had altered binding to palmitoyl-CoA or oleoyl-CoA.
Asunto(s)
Proteínas de Arabidopsis/metabolismo , Arabidopsis/metabolismo , Proteínas Portadoras/metabolismo , Acilcoenzima A/metabolismo , Secuencia de Aminoácidos , Sustitución de Aminoácidos , Arabidopsis/química , Proteínas de Arabidopsis/química , Proteínas Portadoras/química , Escherichia coli , Espacio Extracelular/metabolismo , Histidina , Datos de Secuencia Molecular , Oligopéptidos , Cebollas , Proteínas de Plantas/química , Proteínas de Plantas/metabolismo , Estructura Terciaria de Proteína , Proteínas Recombinantes/metabolismo , Homología Estructural de Proteína , NicotianaRESUMEN
Cytosolic acyl-CoA binding proteins bind long-chain acyl-CoAs and act as intracellular acyl-CoA transporters and pool formers. Recently, we have characterized Arabidopsis thaliana cDNAs encoding novel forms of ACBP, designated ACBP1 and ACBP2, that contain a hydrophobic domain at the N-terminus and show conservation at the acyl-CoA binding domain to cytosolic ACBPs. We have previously demonstrated that ACBP1 is membrane-associated in Arabidopsis. Here, western blot analysis of anti-ACBP2 antibodies on A. thaliana protein showed that ACBP2 is located in the microsome-containing membrane fraction and in the subcellular fraction containing large particles (mitochondria, chloroplasts and peroxisomes), resembling the subcellular localization of ACBP1. To further investigate the subcellular localization of ACBP2, we fused ACBP2 translationally in-frame to GFP. By means of particle gene bombardment, ACBP2-GFP and ACBP1-GFP fusion proteins were observed transiently expressed at the plasma membrane and at the endoplasmic reticulum in onion epidermal cells. GFP fusions with deletion derivatives of ACBPI or ACBP2 lacking the transmembrane domain were impaired in membrane targeting. Our investigations also showed that when the transmembrane domain of ACBP1 or that of ACBP2 was fused with GFP, the fusion protein was targeted to the plasma membrane, thereby establishing their role in membrane targeting. The localization of ACBP1-GFP is consistent with our previous observations using immunoelectron microscopy whereby ACBPI was localized to the plasma membrane and vesicles. We conclude that ACBP2, like ACBP1, is a membrane protein that likely functions in membrane-associated acyl-CoA transfer/metabolism.
Asunto(s)
Proteínas de Arabidopsis/metabolismo , Arabidopsis/metabolismo , Proteínas Portadoras/metabolismo , Proteínas de la Membrana/metabolismo , Proteínas de Plantas/metabolismo , Arabidopsis/genética , Proteínas de Arabidopsis/genética , Western Blotting , Proteínas Portadoras/genética , Membrana Celular/metabolismo , Proteínas Fluorescentes Verdes , Membranas Intracelulares/metabolismo , Proteínas Luminiscentes/genética , Proteínas Luminiscentes/metabolismo , Proteínas de la Membrana/genética , Microscopía Confocal , Cebollas/citología , Cebollas/genética , Cebollas/metabolismo , Proteínas de Plantas/genética , Proteínas Recombinantes de Fusión/genética , Proteínas Recombinantes de Fusión/metabolismoRESUMEN
We have previously shown that SmCP, the gene encoding Solanum melongena cysteine proteinase, is expressed during developmental events associated with programmed cell death (PCD) suggesting its involvement in protein degradation during these events (Xu and Chye, Plant Journal 17 (1999) 321-327). Here, we investigated the regulation of SmCP expression and showed that it is ethylene-inducible and is under circadian control. This circadian rhythm is entrained by light/dark (LD) cycling with peak expression in the late light period, as opposed to that in early light for rbcS, suggesting that protein degradation and photosynthesis are temporally separated by circadian control. Northern blot analysis shows that the pattern of ethylene induction of SmCP is consistent with our previous observation of its significantly increased expression at leaf senescence and fruit ripening when endogenous ethylene is abundant. To further understand SmCP regulation, we have cloned the SmCP promoter and identified a G-box (CACGTG) at -85/-80 by DNase I footprinting analysis of the -221/+17 region. Its specific interaction with nuclear proteins in S. melongena leaves and fruits was confirmed by competitive electrophoretic mobility shift assays using oligonucleotides containing the G-box and mutant derivatives. G-box binding activity was stronger in senescent than young fruits. In circadian-regulated leaves, stronger binding activity coincided with peak circadian expression of SmCP. This correlation between binding activity and expression suggests that G-box binding factors enhance SmCP transcription and that the G-box likely plays a role in circadian regulation of genes affected by LD cycling.