RESUMEN
We report on a measurement of astrophysical tau neutrinos with 9.7 yr of IceCube data. Using convolutional neural networks trained on images derived from simulated events, seven candidate ν_{τ} events were found with visible energies ranging from roughly 20 TeV to 1 PeV and a median expected parent ν_{τ} energy of about 200 TeV. Considering backgrounds from astrophysical and atmospheric neutrinos, and muons from π^{±}/K^{±} decays in atmospheric air showers, we obtain a total estimated background of about 0.5 events, dominated by non-ν_{τ} astrophysical neutrinos. Thus, we rule out the absence of astrophysical ν_{τ} at the 5σ level. The measured astrophysical ν_{τ} flux is consistent with expectations based on previously published IceCube astrophysical neutrino flux measurements and neutrino oscillations.
RESUMEN
Great strides have been made in understanding how membranes and lipid droplets are formed and maintained in land plants, yet much more is to be learned given the complexity of plant lipid metabolism. A complicating factor is the multi-organellar presence of biosynthetic enzymes and unique compositional requirements of different membrane systems. This necessitates a rich network of transporters and transport mechanisms that supply fatty acids, membrane lipids and storage lipids to their final cellular destination. Though we know a large number of the biosynthetic enzymes involved in lipid biosynthesis and a few transport proteins, the regulatory mechanisms, in particular, coordinating expression and/or activity of the majority remain yet to be described. Plants undergoing stress alter their membranes' compositions, and lipids such as phosphatidic acid have been implicated in stress signaling. Additionally, lipid metabolism in chloroplasts supplies precursors for jasmonic acid (JA) biosynthesis, and perturbations in lipid homeostasis has consequences on JA signaling. In this review, several aspects of plant lipid metabolism are discussed that are currently under investigation: cellular transport of lipids, regulation of lipid biosynthesis, roles of lipids in stress signaling, and lastly the structural and oligomeric states of lipid enzymes.
Asunto(s)
Metabolismo de los Lípidos , Plantas/metabolismo , Membrana Celular/metabolismo , Lípidos/biosíntesis , Orgánulos/metabolismoRESUMEN
The photosynthetic (thylakoid) membrane of plants is one of the most extensive biological cell membrane systems found in Nature. It harbours the photosynthetic apparatus, which is essential to life on Earth as carbon dioxide is fixed and atmospheric oxygen released by photosynthesis. Lipid biosynthetic enzymes of different subcellular compartments participate in the biogenesis of the thylakoid membrane system. This process requires the extensive exchange of lipid precursors between the chloroplast and the ER (endoplasmic reticulum). The underlying lipid trafficking phenomena are not yet understood at the mechanistic level, but genetic mutants of the model plant Arabidopsis thaliana with disruptions in lipid trafficking between the ER and the chloroplast have recently become available. Their study has led to the identification of components of the lipid transfer machinery at the inner chloroplast envelope.
Asunto(s)
Arabidopsis/metabolismo , Cloroplastos/metabolismo , Retículo Endoplásmico/metabolismo , Metabolismo de los Lípidos , Transporte Biológico ActivoAsunto(s)
Arabidopsis/genética , Arabidopsis/fisiología , Arabidopsis/citología , Arabidopsis/metabolismo , Evolución Biológica , Tipificación del Cuerpo , Linaje de la Célula , Eliminación de Gen , Regulación de la Expresión Génica de las Plantas , Silenciador del Gen , Genes de Plantas , Genoma de Planta , Genómica , Gravitropismo , Reguladores del Crecimiento de las Plantas/fisiología , Proteínas de Plantas/análisis , Proteínas de Plantas/metabolismo , Estructuras de las Plantas/citología , Estructuras de las Plantas/genética , Estructuras de las Plantas/metabolismo , Estructuras de las Plantas/fisiología , Reproducción , TemperaturaRESUMEN
The galactolipid digalactosyldiacylglycerol (DGGD) is one of the major constituents of thylakoids, accounting for about 25% of polar lipids found in these membranes. Although the presence of DGDG has frequently been correlated with the structural and functional integrity of the photosynthetic apparatus, it is still a matter of debate of what the in-vivo function of DGDG actually might be. To further the understanding of the role of DGDG within the photosynthetic apparatus, experiments were conducted on different Arabidopsis thaliana lines with altered DGDG content. The dgd1 mutant is characterized by a 90% reduction in the DGDG content, resulting in a severe dwarfism during growth. Complementation of the dgd1 mutant with a DGD1 cDNA completely restored the wild-type characteristics, while photosynthesis-related parameters were intermediate in transgenic plants with a partial reduction in DGD1 activity caused by post-transcription gene silencing due to over-expression of a DGD1 cDNA in wild-type plants. These data provide clear evidence for a causal relationship between the DGDG content, and the structure and function of the photosynthetic apparatus. However, a significant DGDG accumulation in the dgd1/pho1 double mutant was without any detectable effect on photosynthetic activity, indicating that the molecular DGDG species synthesized upon phosphate deprivation in leaves cannot substitute for the DGDG species present under normal nutrient supply of plants. It is suggested that depending on the environmental growth conditions different pools of DGDG species exist in plants of which one is not associated with the photosynthetic apparatus.
Asunto(s)
Proteínas de Arabidopsis , Glucolípidos/fisiología , Fosfatos/metabolismo , Fotosíntesis/fisiología , Arabidopsis/genética , Arabidopsis/crecimiento & desarrollo , Arabidopsis/metabolismo , Diglicéridos/metabolismo , Galactolípidos , Galactosiltransferasas/genética , Galactosiltransferasas/metabolismo , Glucolípidos/genética , Glucolípidos/metabolismo , Plantas Modificadas GenéticamenteRESUMEN
Galactolipids make up the bulk of chloroplast lipids. Therefore, the genes involved in the synthesis of the galactolipids monogalactosyldiacylglycerol (MGDG) and digalactosyldiacylglycerol (DGDG) play a critical role in chloroplast development. In this study, we analyzed the subcellular localization of the Arabidopsis DGDG synthase DGD1, which was recently identified by complementation of the Arabidopsis dgd1 mutant. In vitro import experiments demonstrated that DGD1 was targeted to the chloroplast outer envelope in an ATP-independent manner. DGD1 could not be extracted from the membranes by high salt or alkali, suggesting that it is an integral membrane protein. Uptake experiments with truncated versions of DGD1 indicated that the information for targeting and insertion into the outer envelope resides in the N-terminal half of DGD1, but not in the first 33 amino acids. DGD1 apparently does not contain a cleavable signal peptide. Antibodies to Arabidopsis DGD1 detected a 90-kDa protein localized to the chloroplast envelopes of both pea and Arabidopsis. Transformation of DGD1 constructs into cyanobacteria resulted in the expression of active DGDG synthase and demonstrated that DGDG synthesis depends on MGDG lipid, but does not require direct interaction with the plant MGDG synthase.
Asunto(s)
Proteínas de Arabidopsis , Cloroplastos/enzimología , Galactosiltransferasas/metabolismo , Adenosina Trifosfato/metabolismo , Aminoácidos/metabolismo , Arabidopsis , Secuencia de Bases , Western Blotting , Membrana Celular/enzimología , Cartilla de ADN , Galactosiltransferasas/biosíntesis , Galactosiltransferasas/química , Pisum sativum , Transporte de ProteínasRESUMEN
Betaine lipids are ether-linked, nonphosphorous glycerolipids that resemble the more commonly known phosphatidylcholine in overall structure. Betaine lipids are abundant in many eukaryotes such as nonseed plants, algae, fungi, and amoeba. Some of these organisms are entirely devoid of phosphatidylcholine and, instead, contain a betaine lipid such as diacylglyceryl-O-4'-(N,N,N,-trimethyl)homoserine. Recently, this lipid also was discovered in the photosynthetic purple bacterium Rhodobacter sphaeroides where it seems to replace phosphatidylcholine under phosphate-limiting growth conditions. This discovery provided the opportunity to study the biosynthesis of betaine lipids in a bacterial model system. Mutants of R. sphaeroides deficient in the biosynthesis of the betaine lipid were isolated, and two genes essential for this process, btaA and btaB, were identified. It is proposed that btaA encodes an S-adenosylmethionine:diacylglycerol 3-amino-3-carboxypropyl transferase and btaB an S-adenosylmethionine-dependent N-methyltransferase. Both enzymatic activities can account for all reactions of betaine lipid head group biosynthesis. Because the equivalent reactions have been proposed for different eukaryotes, it seems likely that orthologs of btaA/btaB may be present in other betaine lipid-containing organisms.
Asunto(s)
Aminoaciltransferasas/genética , Proteínas Bacterianas , Metiltransferasas/genética , Rhodobacter sphaeroides/enzimología , Triglicéridos/biosíntesis , Aminoaciltransferasas/metabolismo , Secuencia de Bases , Cartilla de ADN , Metiltransferasas/metabolismo , Datos de Secuencia Molecular , Sistemas de Lectura Abierta , Operón , Reacción en Cadena de la Polimerasa , Rhodobacter sphaeroides/genética , Rhodobacter sphaeroides/metabolismoRESUMEN
The sulfolipid sulfoquinovosyldiacylglycerol is a component of plant photosynthetic membranes and represents one of the few naturally occurring sulfonic acids with detergent properties. Sulfolipid biosynthesis involves the transfer of sulfoquinovose, a 6-deoxy-6-sulfoglucose, from UDP-sulfoquinovose to diacylglycerol. The formation of the sulfonic acid precursor, UDP-sulfoquinovose, from UDP-glucose and a sulfur donor is proposed to be catalyzed by the bacterial SQDB proteins or the orthologous plant SQD1 proteins. To investigate the underlying enzymatic mechanism and to elucidate the de novo synthesis of sulfonic acids in biological systems, we developed an in vitro assay for the recombinant SQD1 protein from Arabidopsis thaliana. Among different possible sulfur donors tested, sulfite led to the formation of UDP-sulfoquinovose in the presence of UDP-glucose and SQD1. An SQD1 T145A mutant showed greatly reduced activity. The UDP-sulfoquinovose formed in this assay was identified by co-chromatography with standards and served as substrate for the sulfolipid synthase associated with spinach chloroplast membranes. Approximate K(m) values of 150 microm for UDP-glucose and 10 microm for sulfite were established for SQD1. Based on our results, we propose that SQD1 catalyzes the formation of UDP-sulfoquinovose from UDP-glucose and sulfite, derived from the sulfate reduction pathway in the chloroplast.
Asunto(s)
Arabidopsis/metabolismo , Glucolípidos/metabolismo , Sulfitos/metabolismo , Uridina Difosfato Glucosa/metabolismo , Secuencia de Bases , Cartilla de ADN , Electroforesis en Gel de Poliacrilamida , Glucolípidos/genética , Mutación , Uridina Difosfato Glucosa/análogos & derivadosRESUMEN
To provide a broad analysis of gene expression in developing Arabidopsis seeds, microarrays have been produced that display approximately 2,600 seed-expressed genes. DNA for genes spotted on the arrays were selected from >10,000 clones partially sequenced from a cDNA library of developing seeds. Based on a series of controls, sensitivity of the arrays was estimated at one to two copies of mRNA per cell and cross hybridization was estimated to occur if closely related genes have >70% to 80% sequence identity. These arrays have been hybridized in a series of experiments with probes derived from seeds, leaves, and roots of Arabidopsis. Analysis of expression ratios between the different tissues has allowed the tissue-specific expression patterns of many hundreds of genes to be described for the first time. Approximately 25% of the 2, 600 genes were expressed at ratios > or =2-fold higher in seeds than leaves or roots and 10% at ratios > or =10. Included in this list are a large number of proteins of unknown function, and potential regulatory factors such as protein kinases, phosphatases, and transcription factors. The Arabidopsis arrays were also found to be useful for transcriptional profiling of mRNA isolated from developing oilseed rape (Brassica napus) seeds and expression patterns correlated well between the two species.
Asunto(s)
Arabidopsis/genética , Análisis de Secuencia por Matrices de Oligonucleótidos , Semillas/genética , Brassica/genética , ADN Complementario/genética , Regulación del Desarrollo de la Expresión Génica , Regulación de la Expresión Génica de las PlantasRESUMEN
Large-scale single-pass sequencing of cDNAs from different plants has provided an extensive reservoir for the cloning of genes, the evaluation of tissue-specific gene expression, markers for map-based cloning, and the annotation of genomic sequences. Although as of January 2000 GenBank contained over 220,000 entries of expressed sequence tags (ESTs) from plants, most publicly available plant ESTs are derived from vegetative tissues and relatively few ESTs are specifically derived from developing seeds. However, important morphogenetic processes are exclusively associated with seed and embryo development and the metabolism of seeds is tailored toward the accumulation of economically valuable storage compounds such as oil. Here we describe a new set of ESTs from Arabidopsis, which has been derived from 5- to 13-d-old immature seeds. Close to 28,000 cDNAs have been screened by DNA/DNA hybridization and approximately 10,500 new Arabidopsis ESTs have been generated and analyzed using different bioinformatics tools. Approximately 40% of the ESTs currently have no match in dbEST, suggesting many represent mRNAs derived from genes that are specifically expressed in seeds. Although these data can be mined with many different biological questions in mind, this study emphasizes the import of photosynthate into developing embryos, its conversion into seed oil, and the regulation of this pathway.
Asunto(s)
Arabidopsis/genética , Etiquetas de Secuencia Expresada , Semillas/genética , Arabidopsis/metabolismo , Metabolismo de los Hidratos de Carbono , Proteínas Portadoras/genética , Proteínas Portadoras/metabolismo , Mapeo Cromosómico , ADN Complementario/química , ADN Complementario/genética , Bases de Datos Factuales , Ácidos Grasos/biosíntesis , Regulación del Desarrollo de la Expresión Génica , Regulación de la Expresión Génica de las Plantas , Glucólisis , Vía de Pentosa Fosfato , Fotosíntesis/fisiología , Aceites de Plantas/química , Semillas/metabolismo , Análisis de Secuencia de ADN , Almidón/metabolismoRESUMEN
BACKGROUND: Benign prostate hyperplasia (BPH) and prostate cancer established that disruption of the molecular mechanisms that regulate apoptosis and cell proliferation among the stromal and epithelial cell populations, may underlie the neoplastic development that characterizes the aging gland. This work examined the effects of selected alpha(1)-adrenoceptor (alpha(1)-AR) antagonists (blockers) on cellular dynamics to determine whether induction of apoptosis or inhibition of proliferation could contribute to the overall clinical profile. METHODS: Our efforts were focused on investigating whether alpha(1)-AR antagonists of two different chemical classes affect prostate pathophysiology via mechanisms other than smooth muscle contraction. In in vitro experiments, the two clinically used quinazoline alpha(1)-adrenoceptor antagonists terazosin and doxazosin and the chemically-distinct sulphonamide, tamsulosin, were examined for effects on prostatic tumor growth, by inhibiting cell proliferation and'or inducing apoptosis. RESULTS: Our findings suggest that alpha(1)-AR antagonists, terazosin and doxazosin, suppress prostatic growth by inducing apoptosis in a dose-dependent manner and without affecting cell proliferation. Tamsulosin exerted no effect on prostate cancer cell growth. The apoptotic effect of terazosin and doxazosin appears to be independent of the alpha(1)-adrenoceptor block. CONCLUSIONS: Taken together, our findings demonstrate the ability of the quinazoline alpha-blockers, terazosin and doxazosin, but not the sulphonamide, tamsulosin, to suppress prostate growth by inducing apoptosis among the epithelial cells in the benign and malignant prostate. These studies underwrite the durability of the response seen in long-term studies with terazosin, and suggest the potential of this drug in the treatment of prostate carcinoma.
Asunto(s)
Antagonistas de Receptores Adrenérgicos alfa 1 , Antagonistas Adrenérgicos alfa/farmacología , Apoptosis/efectos de los fármacos , División Celular/efectos de los fármacos , Prazosina/análogos & derivados , Próstata/patología , Enfermedades de la Próstata/tratamiento farmacológico , Biopsia , Doxazosina/farmacología , Humanos , Masculino , Músculo Liso/patología , Prazosina/farmacología , Enfermedades de la Próstata/patología , Hiperplasia Prostática/tratamiento farmacológico , Hiperplasia Prostática/patología , Neoplasias de la Próstata/tratamiento farmacológico , Neoplasias de la Próstata/patología , Células del Estroma/patología , Sulfonamidas/farmacología , TamsulosinaRESUMEN
The galactolipids, mono- and digalactosyldiacylglycerol (DGDG), are the most common nonphosphorous lipids in the biosphere and account for 80% of the membrane lipids found in green plant tissues. These lipids are major constituents of photosynthetic membranes (thylakoids), and a large body of evidence suggests that galactolipids are associated primarily with plastid membranes in seed plants. A null-mutant of Arabidopsis (dgd1), which lacks the DGDG synthase (DGD1) resulting in a 90% reduction in the amount of DGDG under normal growth conditions, accumulated DGDG after phosphate deprivation up to 60% of the amount present in the wild type. This observation suggests the existence of a DGD1-independent pathway of galactolipid biosynthesis. The fatty acid composition of the newly formed DGDG was distinct, showing an enrichment of 16-carbon fatty acids in the C-1 position of the glycerol backbone of DGDG. Roots with their rudimentary plastids accumulated large amounts of DGDG after phosphate deprivation, suggesting that this galactolipid may be located in extraplastidic membranes. Corroborating evidence for this hypothesis was obtained directly by fractionation of subcellular membranes from leaf tissue and indirectly by lipid analysis of the phosphate-deprived fad3 mutant primarily deficient in extraplastidic fatty acid desaturation. The discovery of extraplastidic DGDG biosynthesis induced by phosphate deprivation has revealed a biochemical mechanism for plants to conserve phosphate. Apparently, plants replace phospholipids with nonphosphorous galactolipids if environmental conditions such as phosphate deprivation require this for survival.
Asunto(s)
Proteínas de Arabidopsis , Arabidopsis/metabolismo , Galactosiltransferasas/fisiología , Glucolípidos/biosíntesis , Fosfatos/metabolismo , Arabidopsis/genética , Galactolípidos , Galactosiltransferasas/genética , Fenotipo , Hojas de la Planta/metabolismo , Raíces de Plantas/metabolismo , PlastidiosRESUMEN
Recent evidence from our laboratory has demonstrated that alpha1-adrenoceptor antagonists doxazosin and terazosin induced apoptosis in prostate epithelial and smooth muscle cells in patients with benign prostatic hypertrophy (BPH; J. Urol., 159: 1810-1815, 1998; J. Urol., 161: 2002-2007, 1999). In this study, we investigated the biological action of three alpha1-adrenoceptor antagonists, doxazosin, terazosin, and tamsulosin, against prostate cancer cell growth. The antigrowth effect of the three alpha1-adrenoceptor antagonists was examined in two human prostate cancer cell lines, PC-3 and DU-145, and a prostate smooth muscle cell primary culture, SMC-1, on the basis of: (a) cell viability assay; (b) rate of DNA synthesis; and (c) induction of apoptosis. Our results indicate that treatment of prostate cancer cells with doxazosin or terazosin results in a significant loss of cell viability, via induction of apoptosis in a dose-dependent manner, whereas tamsulosin had no effect on prostate cell growth. Neither doxazosin nor terazosin exerted a significant effect on the rate of cell proliferation in prostate cancer cells. Exposure to phenoxybenzamine, an irreversible inhibitor of alpha1-adrenoceptors, does not abrogate the apoptotic effect of doxazosin or terazosin against human prostate cancer or smooth muscle cells. This suggests that the apoptotic activity of doxazosin and terazosin against prostate cells is independent of their capacity to antagonize alpha1-adrenoceptors. Furthermore, an in vivo efficacy trial demonstrated that doxazosin administration (at tolerated pharmacologically relevant doses) in SCID mice bearing PC-3 prostate cancer xenografts resulted in a significant inhibition of tumor growth. These findings demonstrate the ability of doxazosin and terazosin (but not tamsulosin) to suppress prostate cancer cell growth in vitro and in vivo by inducing apoptosis without affecting cell proliferation. This evidence provides the rationale for targeting both drugs, already in clinical use and with established adverse-effect profiles, against prostatic tumors for the treatment of advanced prostate cancer.
Asunto(s)
Agonistas de Receptores Adrenérgicos alfa 1 , Antagonistas Adrenérgicos alfa/farmacología , Antineoplásicos/farmacología , Apoptosis/efectos de los fármacos , Doxazosina/farmacología , Prazosina/farmacología , Neoplasias de la Próstata/tratamiento farmacológico , Animales , División Celular/efectos de los fármacos , Células Cultivadas , Inhibidores de Crecimiento/farmacología , Humanos , Masculino , Ratones , Ratones SCID , Músculo Liso/citología , Músculo Liso/efectos de los fármacos , Trasplante de Neoplasias , Prazosina/análogos & derivados , Próstata/citología , Próstata/efectos de los fármacos , Neoplasias de la Próstata/metabolismo , Neoplasias de la Próstata/patología , ARN Mensajero/biosíntesis , ARN Mensajero/genética , Receptores Adrenérgicos alfa 1/biosíntesis , Receptores Adrenérgicos alfa 1/genética , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Sulfonamidas/farmacología , Tamsulosina , Trasplante HeterólogoRESUMEN
Large-scale single-pass sequencing of cDNAs prepared from specific plant species or tissues has evolved as an inexpensive and efficient gene-discovery tool that can be used to identify novel cDNAs encoding enzymes of specific plant metabolic pathways. Collections of expressed sequence tags from metabolically active tissues can provide quantitative estimates of gene expression levels and thus are being exploited to unravel plant metabolic and regulatory networks.
Asunto(s)
Etiquetas de Secuencia Expresada , Plantas/metabolismo , Perfilación de la Expresión Génica , Regulación de la Expresión Génica de las Plantas , Plantas/genéticaRESUMEN
The sulfolipid sulfoquinovosyldiacylglycerol is commonly found in the thylakoid membranes of photosynthetic bacteria and plants. While there is a good correlation between the occurrence of sulfolipid and photosynthesis, a number of exceptions are known. Most recently, sulfoquinovosyldiacylglycerol was discovered in the non-photosynthetic, root nodule-forming bacterium Sinorhizobium meliloti. This discovery raised the questions of the phylogenetic origin of genes essential for the biosynthesis of this lipid in S. meliloti and of a function of sulfolipid in root nodule symbiosis. To begin to answer these questions, we isolated and inactivated the sqdB gene of S. meliloti. This gene and two other genes located directly 3' of sqdB are highly similar to the sqdB, sqdC, and sqdD genes known to be essential for sulfolipid biosynthesis in the photosynthetic, purple bacterium Rhodobacter sphaeroides. This observation confirms the close phylogenetic kinship between these two species. Furthermore, the reduced similarity of sqdB to the plant ortholog SQD1 of Arabidopsis thaliana does not support a previous sqd gene transfer from the plant as a consequence of close symbiosis. A sulfolipid-deficient mutant of S. meliloti disrupted in sqdB is capable of inducing functional nodules and does not show an obvious disadvantage under different laboratory culture conditions. Thus far, no specific function can be assigned to bacterial sulfolipid, in either nodule-associated or free-living cells. S. meliloti contains a rich set of polar membrane lipids some of which, including sulfolipid, may become critical only under growth conditions that still need to be discovered.
Asunto(s)
Proteínas de Arabidopsis , Proteínas Bacterianas/genética , Glucolípidos/biosíntesis , Sinorhizobium meliloti/genética , Simbiosis , Proteínas Bacterianas/metabolismo , Southern Blotting , Cromatografía en Capa Delgada , Glucolípidos/genética , Medicago sativa/microbiología , Medicago sativa/fisiología , Datos de Secuencia Molecular , Mutagénesis Insercional , Proteínas de Plantas/genética , Raíces de Plantas/microbiología , Raíces de Plantas/fisiología , Sinorhizobium meliloti/metabolismoRESUMEN
The lipid monogalactosyl diacylglycerol (MGD) is a major structural component of photosynthetic membranes in chloroplasts. Its formation is catalyzed by the enzyme MGD synthase. In many plants, MGD derives from two different biosynthetic pathways: the prokaryotic pathway, which operates entirely within the plastid, and the eukaryotic pathway, which involves steps in the endoplasmic reticulum. Here, we describe the identification and characterization of an Arabidopsis mutant with a defective MGD synthase gene (MGD1). The mutant was identified in a screen of T-DNA lines for individuals with defects in chloroplast biogenesis. It has a yellow-green phenotype that correlates with a approximately 50% deficiency in total chlorophyll per plant. A single T-DNA insertion is located adjacent to the transcription initiation site of the MGD1 gene, and the abundance of MGD1 mRNA is reduced by 75% compared with wild type. Correlation between steady-state MGD1 transcript levels and MGD synthase activity (also reduced by 75% in mgd1) suggests that MGD1 is the most important MGD synthase in green tissues. The amount of MGD in mutant leaves is reduced by 42% compared with wild type. MGD from the mutant contains 23% less 16:3 fatty acid and 10% more 18:3 fatty acid. Because 16:3 is a characteristic feature of MGD from the prokaryotic pathway, it is possible that MGD1 operates with some preference in the prokaryotic pathway. Finally, the MGD-deficiency of mgd1 is correlated with striking defects in chloroplast ultrastructure, strongly suggesting a unique role for MGD in the structural organization of plastidic membranes.
Asunto(s)
Arabidopsis/genética , Cloroplastos/genética , Galactosiltransferasas/genética , Glucolípidos/biosíntesis , Secuencia de Aminoácidos , Arabidopsis/enzimología , Cloroplastos/enzimología , Cloroplastos/ultraestructura , Galactolípidos , Genes de Plantas , Isoenzimas/genética , Datos de Secuencia Molecular , Mutación , Fenotipo , Homología de Secuencia de AminoácidoRESUMEN
The sulfolipid sulfoquinovosyldiacylglycerol is present in the photosynthetic membranes of plants and many photosynthetic bacteria. A novel gene, sqdX, essential for sulfolipid biosynthesis in the cyanobacterium Synechococcus sp. strain PCC7942 is proposed to encode the cyanobacterial sulfolipid synthase catalyzing the last reaction of the pathway.
Asunto(s)
Proteínas de Arabidopsis , Cianobacterias/genética , Glucolípidos/biosíntesis , Proteínas de Plantas/genética , Proteínas Bacterianas/genética , Cianobacterias/enzimología , Datos de Secuencia Molecular , Sistemas de Lectura Abierta , FotosíntesisRESUMEN
For over 25 years there has been uncertainty over the pathway from CO(2) to acetyl-CoA in chloroplasts. On the one hand, free acetate is the most effective substrate for fatty acid synthesis by isolated chloroplasts, and free acetate concentrations reported in leaf tissue (0.1-1 mM) appear adequate to saturate fatty acid synthase. On the other hand, a clear mechanism to generate sufficient free acetate for fatty acid synthesis is not established and direct production of acetyl-CoA from pyruvate by a plastid pyruvate dehydrogenase seems a more simple and direct path. We have re-examined this question and attempted to distinguish between the alternatives. The kinetics of (13)CO(2) and (14)CO(2) movement into fatty acids and the absolute rate of fatty acid synthesis in leaves was determined in light and dark. Because administered (14)C appears in fatty acids within < 2-3 min our results are inconsistent with a large pool of free acetate as an intermediate in leaf fatty acid synthesis. In addition, these studies provide an estimate of the turnover rate of fatty acid in leaves. Studies similar to the above are more complex in seeds, and some questions about the regulation of plant lipid metabolism seem difficult to solve using conventional biochemical or molecular approaches. For example, we have little understanding of why or how some seeds produce >50% oil whereas other seeds store largely carbohydrate or protein. Major control over complex plant biochemical pathways may only become possible by understanding regulatory networks which provide 'global' control over these pathways. To begin to discover such networks and provide a broad analysis of gene expression in developing oilseeds, we have produced microarrays that display approx. 5000 seed-expressed Arabidopsis genes. Sensitivity of the arrays was 1-2 copies of mRNA/cell. The arrays have been hybridized with probes derived from seeds, leaves and roots, and analysis of expression ratios between the different tissues has allowed the tissue-specific expression patterns of many hundreds of genes to be described for the first time. Approx. 10% of the genes were expressed at ratios > or = 10-fold higher in seeds than in leaves or roots. Included in this list are a large number of proteins of unknown function, and potential regulatory factors such as protein kinases, phosphatases and transcription factors. The arrays were also found to be useful for analysis of Brassica seeds.
Asunto(s)
Dióxido de Carbono/metabolismo , Cloroplastos/metabolismo , Ácidos Grasos/biosíntesis , Genómica , Plantas/genética , Plantas/metabolismo , Acetilcoenzima A/metabolismoRESUMEN
To explore the role of digalactosyldiacylglycerol (DGDG) in plants the dgd1 mutant of Arabidopsis thaliana was grown in the presence and absence of inorganic phosphate. Phosphate deficiency in the dgd1 mutant causes a strong decrease in all phospholipids accompanied by an increase in DGDG and sulpholipid. Moreover, a significant DGDG accumulation was found in roots upon phosphate deprivation as well. Our data indicate that DGDG accumulation upon phosphate deprivation is due to the activation of a specific eukaryotic dgd1-independent biosynthetic pathway. We propose that DGDG may substitute for phosphatidylcholine upon phosphate deprivation.
Asunto(s)
Arabidopsis/metabolismo , Galactolípidos , Glucolípidos/metabolismo , Fosfatos/metabolismo , Fosfatidilcolinas/metabolismo , Fosfolípidos/metabolismo , Arabidopsis/genética , Ácidos Grasos/análisis , Glucolípidos/genética , Fosfolípidos/química , Hojas de la Planta/metabolismo , Raíces de Plantas/metabolismoRESUMEN
Diacylglycerol acyltransferase (DGAT, EC 2.3.1.20) is a membrane enzyme that drives the final step in the formation of oils using diacylglycerol (DAG) and acyl-CoA to yield triacylglycerol (TAG). We identified a putative plant DGAT gene (TRIACYLGLYCEROL1: TAG1) and demonstrated its function by the cloning of two mutated alleles, designated AS11 (tag1-1) and ABX45 (tag1-2). One allele, AS11, has been previously characterised at the biochemical level. Mutant seeds contained less oil with a modified fatty acid profile and have reduced germination rates compared to wild-type controls. The TAG1 cDNA encodes for a 520-aa protein that possesses multiple putative transmembrane domains and shows 70 % similarity to a human DGAT cDNA.