RESUMEN
The conversion of etioplasts into chloroplasts in germinating cotyledons is a crucial transition for higher plants, enabling photoautotrophic growth upon illumination. Tight coordination of chlorophyll biosynthesis and photosynthetic complex assembly is critical for this process. ORANGE (OR), a DnaJ-like zinc finger domain-containing protein, was reported to trigger the biogenesis of carotenoid-accumulating plastids by promoting carotenoid biosynthesis and sequestration. Both nuclear and plastidic localizations of OR have been observed. Here, we show that Arabidopsis (Arabidopsis thaliana) OR physically interacts with the transcription factor TCP14 in the nucleus and represses its transactivation activity. Through this interaction, the nucleus-localized OR negatively regulates expression of EARLY LIGHT-INDUCIBLE PROTEINS (ELIPs), reduces chlorophyll biosynthesis, and delays development of thylakoid membranes in the plastids of germinating cotyledons. Nuclear abundance of OR decreased upon illumination. Together with an accumulation of TCP14 in the nucleus, this derepresses chloroplast biogenesis during de-etiolation. TCP14 is epistatic to OR and expression of ELIPs is directly regulated by the binding of TCP14 to Up1 elements in the ELIP promoter regions. Our results demonstrate that the interaction between OR and TCP14 in the nucleus leads to repression of chloroplast biogenesis in etiolated seedlings and provide new insights into the regulation of early chloroplast development.plantcell;31/12/2996/FX1F1fx1.
Asunto(s)
Proteínas de Arabidopsis/metabolismo , Arabidopsis/metabolismo , Clorofila/biosíntesis , Cloroplastos/metabolismo , Cotiledón/metabolismo , Proteínas del Choque Térmico HSP40/metabolismo , Factores de Transcripción/metabolismo , Arabidopsis/genética , Proteínas de Arabidopsis/genética , Proteínas de Arabidopsis/efectos de la radiación , Núcleo Celular/metabolismo , Núcleo Celular/efectos de la radiación , Cotiledón/genética , Etiolado , Regulación de la Expresión Génica de las Plantas/genética , Germinación , Proteínas del Choque Térmico HSP40/genética , Proteínas del Choque Térmico HSP40/efectos de la radiación , Iluminación , Plastidios/metabolismo , Regiones Promotoras Genéticas , Unión Proteica , Plantones/metabolismo , Tilacoides/metabolismo , Factores de Transcripción/genética , Regulación hacia ArribaRESUMEN
Carotenoids are essential phytonutrients for the human body. Higher plants usually synthesize and accumulate carotenoids in their leaves, flowers, and fruits. Most carotenoids have either two ß-rings on both ends or ß- and ε-rings separately on two ends of their molecules and are synthesized from the acyclic lycopene as the precursor. Lycopene ß- and ε-cyclases (LCYB and LCYE, respectively) catalyze the ß- and ε-cyclization of lycopene, respectively, and regulate the metabolic flux from lycopene to its downstream ß,ß-branches (by LCYB alone) and ß,ε-branches (by LCYE and LCYB). In this study, we identified and characterized genes for two LCYBs (CaLCYB1 and CaLCYB2), one LCYE (CaLCYE1), and a capsanthin/capsorubin synthase (CaCCS1) which is also able to ß-cyclize lycopene from the red pepper ( Capsicum annuum var. conoides) genome. By quantifying transcript abundances of these genes and contents of different carotenoid components in ripening fruits, we observed a correlation between the induction of both CaLCYBs and the accumulation of carotenoids of the ß,ß-branch during ripening. Although capsanthin was accumulated in ripened fruits, our quantification demonstrated a strong induction of CaCCS1 at the breaker stage, together with the simultaneous repression of CaLCYE1 and the decrease of lutein content, suggesting the involvement of CaCCS1 in competing against CaLCYE1 for synthesizing carotenoids of the ß,ß-branch. Our results provide important information for future metabolic engineering studies to manipulate carotenoid biosynthesis and accumulation in fruits.
Asunto(s)
Capsicum/metabolismo , Carotenoides/metabolismo , Licopeno/química , Licopeno/metabolismo , Capsicum/genética , Capsicum/crecimiento & desarrollo , Ciclización , Frutas/genética , Frutas/crecimiento & desarrollo , Frutas/metabolismo , Regulación de la Expresión Génica de las Plantas , Liasas Intramoleculares/genética , Liasas Intramoleculares/metabolismo , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismoRESUMEN
Chlorophylls and carotenoids not only function in photosynthesis and photoprotection but are also involved in the assembly of thylakoid membranes and the stabilization of apoproteins in photosystems. In this study, we identified a nuclear gene required for chlorophyll and carotenoid metabolism, namely, DWARF AND YELLOW 1 (DY1). Growth of the loss-of-function dy1 mutant was severely retarded, and the seedlings of this mutant accumulated significantly less amounts of both chlorophylls and carotenoids in cotyledons and rosette leaves, although genes related to pigment metabolism did not show corresponding fluctuation at the transcriptional level. In chloroplasts of the dy1 leaves, thylakoids were loosely packed into grana. The dy1 mutant also possessed severely impaired photosynthetic and photoprotective abilities. DY1 encodes a chloroplast stroma protein that is highly conserved in vascular plants. Our results demonstrated that after the full-length DY1 (53 kDa) was imported into the chloroplast and its N-terminal transit peptide was processed, the C-terminal end of this premature DY1 (42 kDa) was also removed during the maturation of rosette leaves, resulting in a 24-kDa mature peptide. Our blue native PAGE and Western blot analyses showed the presence of both premature and mature forms of DY1 in protein complexes. The involvement of DY1 in chloroplast development is discussed.
RESUMEN
The biosynthesis of chlorophylls and carotenoids and the assembly of thylakoid membranes are critical for the photoautotrophic growth of plants. Different factors are involved in these two processes. In recent years, members of the DnaJ-like zinc finger domain proteins have been found to take part in the biogenesis and/or the maintenance of plastids. One member of this family of proteins, PSA2, was recently found to localize to the thylakoid lumen and regulate the accumulation of photosystem I. In this study, we report that the silencing of PSA2 in Arabidopsis thaliana resulted in variegated leaves and retarded growth. Although both chlorophylls and total carotenoids decreased in the psa2 mutant, violaxanthin, and zeaxanthin accumulated in the mutant seedlings grown under growth condition. Lower levels of non-photochemical quenching and electron transport rate were also found in the psa2 mutant seedlings under growth condition compared with those of the wild-type plants, indicating an impaired capability to acclimate to normal light irradiance when PSA2 was silenced. Moreover, we also observed an abnormal assembly of grana thylakoids and poorly developed stroma thylakoids in psa2 chloroplasts. Taken together, our results demonstrate that PSA2 is a member of the DnaJ-like zinc finger domain protein family that affects light acclimation and chloroplast development.
RESUMEN
Two series of thiazolidinone derivatives designing for potential EGFR and HER-2 kinase inhibitors have been discovered. Some of them exhibited significant EGFR and HER-2 inhibitory activity. Compound 2-(2-(5-bromo-2-hydroxybenzylidene)hydrazinyl)thiazol-4(5H)-one (12) displayed the most potent inhibitory activity (IC(50)=0.09 microM for EGFR and IC(50)=0.42 microM for HER-2), comparable to the positive control erlotinib. Docking simulation was performed to position compound 12 into the EGFR active site to determine the probable binding model. Antiproliferative assay results indicating that some of the thiazolidinone derivatives own high antiproliferative activity against MCF-7. Compound 12 with potent inhibitory activity in tumor growth inhibition would be a potential anticancer agent.
Asunto(s)
Antineoplásicos/química , Antineoplásicos/farmacología , Receptores ErbB/antagonistas & inhibidores , Receptor ErbB-2/antagonistas & inhibidores , Tiazolidinas/química , Tiazolidinas/farmacología , Antineoplásicos/síntesis química , Línea Celular Tumoral , Proliferación Celular/efectos de los fármacos , Ensayos de Selección de Medicamentos Antitumorales , Receptores ErbB/química , Receptores ErbB/metabolismo , Humanos , Modelos Moleculares , Neoplasias/tratamiento farmacológico , Unión Proteica , Inhibidores de Proteínas Quinasas/síntesis química , Inhibidores de Proteínas Quinasas/química , Inhibidores de Proteínas Quinasas/farmacología , Receptor ErbB-2/metabolismo , Relación Estructura-Actividad , Tiazolidinas/síntesis químicaRESUMEN
Two series of novel N-benzyl-N-(X-2-hydroxybenzyl)-N'-phenylureas and thioureas (1a-18a; 1b-18b) as potential EGFR and HER-2 kinase inhibitors have been discovered. These compounds displayed good EGFR and HER-2 inhibitory activity and the SARs are also been studied. Especially compound 7b demonstrated significant EGFR and HER-2 inhibitory activity (IC(50)=0.08 microM for EGFR and IC(50)=0.35 microM for HER-2). Docking simulation was performed to position compound 7b into the EGFR active site to determine the probable binding conformation and antiproliferative assay results indicating that these series of urea and thioureas own high antiproliferative activity against MCF-7. Above all, thiourea 7b would be a potential anticancer agent deserves further research.
Asunto(s)
Antineoplásicos/química , Antineoplásicos/farmacología , Compuestos de Fenilurea/química , Compuestos de Fenilurea/farmacología , Tiourea/química , Tiourea/farmacología , Antineoplásicos/síntesis química , Línea Celular Tumoral , Proliferación Celular/efectos de los fármacos , Ensayos de Selección de Medicamentos Antitumorales , Receptores ErbB/antagonistas & inhibidores , Receptores ErbB/química , Receptores ErbB/metabolismo , Humanos , Modelos Moleculares , Neoplasias/tratamiento farmacológico , Compuestos de Fenilurea/síntesis química , Unión Proteica , Conformación Proteica , Receptor ErbB-2/antagonistas & inhibidores , Receptor ErbB-2/metabolismo , Relación Estructura-Actividad , Tiourea/síntesis químicaRESUMEN
A series of amide-coupled benzoic nitrogen mustard derivatives as potential EGFR and HER-2 kinase inhibitors were synthesized and reported for the first time. Some of them exhibited significant EGFR and HER-2 inhibitory activity. Of all the studied compounds, compounds 5b and 5t exhibited the most potent inhibitory activity, which was comparable to the positive control erlotinib. Docking simulation was performed to position compounds 5b and 5t into the EGFR active site to determine the probable binding model. Antiproliferative assay results indicated that some of the benzoic nitrogen mustard derivatives possessed high antiproliferative activity against MCF-7. In particular, compounds 5b and 5t with potent inhibitory activity in tumor growth inhibition may function as potential antitumor agents.
Asunto(s)
Amidas/farmacología , Antineoplásicos/farmacología , Compuestos de Mostaza Nitrogenada/farmacología , Inhibidores de Proteínas Quinasas/farmacología , Amidas/síntesis química , Amidas/química , Antineoplásicos/síntesis química , Antineoplásicos/química , Línea Celular Tumoral , Proliferación Celular/efectos de los fármacos , Ensayos de Selección de Medicamentos Antitumorales , Receptores ErbB/antagonistas & inhibidores , Humanos , Modelos Moleculares , Simulación de Dinámica Molecular , Estructura Molecular , Compuestos de Mostaza Nitrogenada/síntesis química , Compuestos de Mostaza Nitrogenada/química , Inhibidores de Proteínas Quinasas/síntesis química , Inhibidores de Proteínas Quinasas/química , Receptor ErbB-2/antagonistas & inhibidores , Estereoisomerismo , Relación Estructura-ActividadRESUMEN
Four nutrient levels, i.e., 0.5 mg N x L(-1) and 0.1 mg P x L(-1) (I), 1.5 mg N x L(-1) and 0.3 mg P x L(-1) (II), 4.5 mg N x L(-1) and 0.9 mg P x L(-1) (III), and 13.5 mg N x L(-1) and 2.7 mg P x L(-1) (IV), were installed to study the effects of water body's nutrient level, plant species combination, and their interactions on the biomass accumulation and allocation of invasive floating species Eichhornia crassipes, native rooted leaf-floating species Jussiaea stipulacea, and submerged plant Vallisneria spiralis. The total, root, stem, and leaf biomass of E. crassipes and J. stipulacea, either in monoculture or in mixed-culture, increased with increasing water body's nutrient level, their total biomass in treatments III and IV being averagely 54.47% and 102.63% higher than that in treatments I and II, respectively. Under different plant species combination, the total, root, stem, and leaf biomass of V. spiralis showed a declining trend with the increase of nutrient level, and the total biomass of V. spiralis in treatments III and IV was averagely 45.88% lower than that in treatments I and II. The results of two-way ANOVA showed that water body's nutrient level had significant positive effects on the biomass of E. crassipes and J. stipulacea but negative effects on that of V. spiralis, and the effects of plant species combination varied with target plant species.