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971.
Lingling Feng Hui Li Jingmei Jiao Ding Li Li Zhou Jian Wan Yangsheng Li 《Journal of Plant Biology》2009,52(5):382-394
Rice cultivar zhonghua11 (Oryza sativa L. ssp. japonica) plants with decreased sedoheptulose-1, 7-bisphosphatase (SBPase) were obtained by transformation with the rice SBPase antisense
gene under the control of the maize ubiquitin promoter. The transgenic and wild-type plants were grown at different nitrogen
levels (0.1, 1, or 10 mM NH4NO3). Growth rates of the seedlings were measured by the changes in dry weight, and the photosynthetic carbon reduction activities
and the potential efficiency of photosystem II were measured by CO2 assimilation and F
v/F
m, respectively. At low N, there are strong effects on growth and photosynthesis when SBPase was reduced by genetic manipulation.
Decreased SBPase activity led to a decrease in the amount of starch accumulated in the leaves at all N levels and the decrease
was much more prominent in low N than that in high N, but the starch allocation between shoot and root was unaltered. The
analysis of chlorophyll fluorescence and SBPase activity indicated that the decrease of growth and photosynthesis at different
N levels were not related to the function of PSII but to the activity of SBPase. Western blot analysis showed the content
of SBPase in thylakoid membranes was much more than in the stroma fractions in transgenic plants at low N. Results suggested
that low N in addition to a 34% decrease in SBPase activity is sufficient to diminish photosynthesis and limit biomass production.
Decreased SBPase activity may reduce the N use efficiency of photosynthesis and growth and alter biomass allocation. 相似文献
972.
Hui Yang Lixin Wan Fuchun Song Mengxi Wang Youguo Huang 《The international journal of biochemistry & cell biology》2009,41(7):1495-1501
Interaction between GAP-43 (growth associated protein-43) and Gαo (alpha subunit of Go protein) influences the signal transduction pathways leading to differentiation of neural cells. GAP-43 is known to increase guanine nucleotide exchange by Gαo, which is a major component of neuronal growth cone membranes. However, it is not clear whether GAP-43 stimulation is related to the Gαo palmitoylation or the conversion of Gαo from oligmers to monomers, which was shown to be a necessary regulatory factor in GDP/GTP exchange of Gαo. Here we expressed and purified GAP-43, GST-GAP-43 and Gαo proteins, detected their stimulatory effect on [35S]-GTPγS binding of Gαo. It was found that the EC50 of both GAP-43 and GST-GAP-43 activation were tenfold lower in case of depalmitoylated Gαo than palmitoylated Gαo. Non-denaturing gel electrophoresis and p-PDM cross-linking analysis revealed that addition of GST-GAP-43 induced disassociation of depalmitoylated Gαo from oligomers to monomers, but did not influence the oligomeric state of palmitoylated Gαo, which suggests that palmitoylation is a key regulatory factor in GAP-43 stimulation on Gαo. These results indicated the interaction of GAP-43 and Gαo could accelerate conversion of depalmitoylated Gαo but not palmitoylated Gαo from oligomers to monomers, so as to increase the GTPγS binding activity of Gαo. Results here provide new evidence about how signaling protein palmitoylation is involved in the G-protein-coupled signal transduction cascade, and give a useful clue on the participation of GAP-43 in G-protein cycle by its preferential activation of depalmitoylated Gαo. 相似文献
973.
Markus Islinger Ka Wan Li Jürgen Seitz Alfred Völkl Georg H. Lüers 《Traffic (Copenhagen, Denmark)》2009,10(11):1711-1721
Most newly synthesized peroxisomal proteins are imported in a receptor-mediated fashion, depending on the interaction of a peroxisomal targeting signal (PTS) with its cognate targeting receptor Pex5 or Pex7 located in the cytoplasm. Apart from this classic mechanism, heterologous protein complexes that have been proposed more than a decade ago are also to be imported into peroxisomes. However, it remains still unclear if this so-called piggyback import is of physiological relevance in mammals. Here, we show that Cu/Zn superoxide dismutase 1 (SOD1), an enzyme without an endogenous PTS, is targeted to peroxisomes using its physiological interaction partner 'copper chaperone of SOD1' (CCS) as a shuttle. Both proteins have been identified as peroxisomal constituents by 2D-liquid chromatography mass spectrometry of isolated rat liver peroxisomes. Yet, while a major fraction of CCS was imported into peroxisomes in a PTS1-dependent fashion in CHO cells, overexpressed SOD1 remained in the cytoplasm. However, increasing the concentrations of both CCS and SOD1 led to an enrichment of SOD1 in peroxisomes. In contrast, CCS-mediated SOD1 import into peroxisomes was abolished by deletion of the SOD domain of CCS, which is required for heterodimer formation. SOD1/CCS co-import is the first demonstration of a physiologically relevant piggyback import into mammalian peroxisomes. 相似文献
974.
Non-Additive Effects of Water and Nitrogen Addition on Ecosystem Carbon Exchange in a Temperate Steppe 总被引:1,自引:0,他引:1
Shuli Niu Haijun Yang Zhe Zhang Mingyu Wu Qi Lu Linghao Li Xingguo Han Shiqiang Wan 《Ecosystems》2009,12(6):915-926
Changes in precipitation and nitrogen (N) deposition can influence ecosystem carbon (C) cycling and budget in terrestrial
biomes, with consequent feedbacks to climate change. However, little is known about the main and interactive effects of water
and N additions on net ecosystem C exchange (NEE). In a temperate steppe of northern China, a field-manipulated experiment
was conducted to evaluate the responses of NEE and its components to improve N and water availability from 2005 to 2008. The
results showed that both water and N additions stimulated gross ecosystem productivity (GEP), ecosystem respiration (ER),
and NEE. Water addition increased GEP by 17%, ER by 24%, and NEE by 11% during the experimental period, whereas N addition
increased GEP by 17%, ER by 16%, and NEE by 19%. The main effects of both water and N additions changed with time, with the
strongest water stimulation in the dry year and a diminishing N stimulation over time. When water and N were added in combination,
there were non-additive effects of water and N on ecosystem C fluxes, which could be explained by the changes in species composition
and the shifts of limiting resources from belowground (water or N) to aboveground (light). The positive water and N additions
effects indicate that increasing precipitation and N deposition in the future will favor C sequestration in the temperate
steppe. The non-additive effects of water and N on ecosystem C fluxes suggest that multifactor experiments are better able
to capture complex interactive processes, thus improving model simulations and projections. 相似文献
975.
Alessandra Pesce Liesbet Thijs Marco Nardini Lorenza Sisinni Louise Gourlay Massimiliano Coletta Xuehua Wan Paolo Ascenzi Martino Bolognesi 《Journal of molecular biology》2009,386(1):246-35534
Among heme-based sensors, recent phylogenomic and sequence analyses have identified 34 globin coupled sensors (GCS), to which an aerotactic or gene-regulating function has been tentatively ascribed. Here, the structural and biochemical characterization of the globin domain of the GCS from Geobacter sulfurreducens (GsGCS162) is reported. A combination of X-ray crystallography (crystal structure at 1.5 Å resolution), UV-vis and resonance Raman spectroscopy reveals the ferric GsGCS162 as an example of bis-histidyl hexa-coordinated GCS. In contrast to the known hexa-coordinated globins, the distal heme-coordination in ferric GsGCS162 is provided by a His residue unexpectedly located at the E11 topological site. Furthermore, UV-vis and resonance Raman spectroscopy indicated that ferrous deoxygenated GsGCS162 is a penta-/hexa-coordinated mixture, and the heme hexa-to-penta-coordination transition does not represent a rate-limiting step for carbonylation kinetics. Lastly, electron paramagnetic resonance indicates that ferrous nitrosylated GsGCS162 is a penta-coordinated species, where the proximal HisF8-Fe bond is severed. 相似文献
976.
977.
978.
上海地区番茄黄化曲叶病毒病的鉴定及嫁接接种法研究 总被引:4,自引:0,他引:4
番茄黄化曲叶病毒(tomato yellow leafcurl virus,TYLCV)是一种由烟粉虱(Bemisia tabaci)和嫁接传播的双生病毒,在热带、亚热带地区给番茄生产造成严重威胁.根据番茄黄化曲叶病毒的保守序列设计一对引物,运用PCR技术从上海地区的感病番茄中扩增出一条575bp的特异带,而健康植株无此带.测序表明该序列与番茄黄化曲叶病毒具有极高的同源性(97%~99%).将健康接穗嫁接到感染番茄黄化曲叶病毒的番茄砧木上,间隔15 d和30 d,分别提取接穗的DNA,并用PCR法检测病毒,发现嫁接15 d后在部分接穗中检测到TYLCV病毒,嫁接30 d后在所有的接穗中均检测到病毒,因此,嫁接法可以作为番茄黄化曲叶病毒病的接种鉴定方法. 相似文献
979.
The Oxa1 protein plays a central role in facilitating the cotranslational insertion of the nascent polypeptide chains into the mitochondrial inner membrane. Mitochondrially encoded proteins are synthesized on matrix-localized ribosomes which are tethered to the inner membrane and in physical association with the Oxa1 protein. In the present study we used a chemical cross-linking approach to map the Saccharomyces cerevisiae Oxa1-ribosome interface, and we demonstrate here a close association of Oxa1 and the large ribosomal subunit protein, MrpL40. Evidence to indicate that a close physical and functional relationship exists between MrpL40 and another large ribosomal protein, the Mrp20/L23 protein, is also provided. MrpL40 shares sequence features with the bacterial ribosomal protein L24, which like Mrp20/L23 is known to be located adjacent to the ribosomal polypeptide exit site. We propose therefore that MrpL40 represents the Saccharomyces cerevisiae L24 homolog. MrpL40, like many mitochondrial ribosomal proteins, contains a C-terminal extension region that bears no similarity to the bacterial counterpart. We show that this C-terminal mitochondria-specific region is important for MrpL40''s ability to support the synthesis of the correct complement of mitochondrially encoded proteins and their subsequent assembly into oxidative phosphorylation complexes.The mitochondrial genome encodes a small, but important, number of proteins (8). These proteins are predominantly essential components of the mitochondrial oxidative phosphorylation (OXPHOS) machinery. In the yeast Saccharomyces cerevisiae the proteins encoded by the mitochondrial DNA (mtDNA) include cytochrome c oxidase subunits Cox1, Cox2, and Cox3, cytochrome b of the cytochrome bc1 complex, F1Fo-ATP synthase subunits Atp6, Atp8, and Atp9, and the small ribosomal subunit component Var1. With the exception of Var1, these mitochondrially encoded proteins are integral membrane proteins which become inserted into the inner membrane during their synthesis on mitochondrial ribosomes tethered to the inner membrane (11, 19, 29, 32, 34). The cotranslational membrane insertion of these proteins is achieved by maintaining a close physical association of the ribosomes to the inner membrane at sites where the insertion machinery exists (19, 31, 32).Oxa1 is an inner membrane protein that forms a central component of the insertion machinery, whose presence is required for the cotranslational membrane insertion of the mitochondrially encoded proteins (4-6, 15-17). The Oxa1 protein has been shown to physically associate with the ribosomes and more specifically with the large ribosomal subunit. Matrix-exposed elements of the Oxa1 protein, such as its hydrophilic C-terminal tail, support this Oxa1-ribosome interaction (19, 32). Furthermore, in intact mitochondria we have previously demonstrated that Oxa1 can be chemically cross-linked to Mrp20, a component of the large ribosomal subunit (19). Mrp20 is homologous to the bacterial ribosomal protein L23, a component known from the structural analysis of the ribosomes to be located next to the polypeptide exit site of the large ribosomal subunit (3, 10, 23, 27, 30). Thus, it was concluded that Oxa1, the site of membrane insertion into the inner membrane, exists in close physical proximity to the large ribosomal subunit and specifically to that region of the ribosomes where the nascent chain emerges. This close physical relationship between ribosomal components and the Oxa1 insertion site has been proposed to support a tight coordination between the protein translation and membrane insertion events (19, 31, 32). Given the strong hydrophobicity of the OXPHOS complex subunits which are encoded by the mitochondrial DNA and synthesized by these ribosomes, a close coupling of the translation and insertion events is proposed to ensure that the hydrophobic nascent chains are directly inserted into the membrane during their synthesis. The exposure of hydrophobic nascent chains to the hydrophilic matrix space may promote their aggregation and thus incompetency for subsequence membrane insertion.In bacteria, the L23 protein has been implicated to play a direct role in the cotranslational insertion of proteins into the membrane (7, 13, 24, 33). Thus, it is possible that proteins adjacent to the polypeptide exit site of mitochondrial ribosomes may be directly involved in targeting ribosomes to specific regions of the inner membrane where the membrane insertion and subsequent assembly events occur. The mitochondrial ribosomes resemble their prokaryotic ancestors in some respects, e.g., antibiotic sensitivity, but they differ in a number of important ways (1, 12, 22, 30). In general, the protein content of the mitochondrial ribosomes is greater than their bacterial counterparts. This increase in protein content is largely attributed to the fact that the mitochondrial ribosomal proteins are larger in size than their bacterial homologs. Over the course of evolution, many of the mitochondrial ribosomal proteins have acquired novel extensions, new domains, in addition to their bacterial homology domains. These acquired extensions not only include N-terminal (often cleavable) signals to target these proteins (nuclear encoded) to the mitochondria but also in many instances large C-terminal extensions, which are unique to the mitochondrial ribosomal proteins and have thus been termed “mitospecific domains” (12, 30). Largely uncharacterized, the functional relevance of these various mitospecific domains of the ribosomal proteins remains unknown. It is speculated that some (or all) of these mitospecific domains serve to ensure that the ribosome becomes assembled and is translationally active while bound to the inner membrane surface.In the present study we sought to further characterize the interaction of the mitochondrial ribosome with the Oxa1 protein. We show here that MrpL40, a large ribosomal subunit component, is physically close to both the Mrp20 and Oxa1 proteins, demonstrating the proximity of MrpL40 to both the ribosomal polypeptide exit site and the Oxa1 membrane insertion site. MrpL40 contains a large C-terminal mitospecific domain, which includes a predicted α-helical region at its extreme C-terminal end. The results presented here highlight that the integrity of this domain of MrpL40 is crucial to ensure ribosome translational fidelity and subsequent OXPHOS complex assembly. 相似文献
980.