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1.
Vibrio cholerae is autochthonous to various aquatic niches and is the etiological agent of the life-threatening diarrheal disease cholera. The persistence of V. cholerae in natural habitats is a crucial factor in the epidemiology of cholera. In contrast to the well-studied V. cholerae-chitin connection, scarce information is available about the factors employed by the bacteria for the interaction with collagens. Collagens might serve as biologically relevant substrates, because they are the most abundant protein constituents of metazoan tissues and V. cholerae has been identified in association with invertebrate and vertebrate marine animals, as well as in a benthic zone of the ocean where organic matter, including collagens, accumulates. Here, we describe the characterization of the V. cholerae putative collagenase, VchC, encoded by open reading frame VC1650 and belonging to the subfamily M9A peptidases. Our studies demonstrate that VchC is an extracellular collagenase degrading native type I collagen of fish and mammalian origin. Alteration of the predicted catalytic residues coordinating zinc ions completely abolished the protein enzymatic activity but did not affect the translocation of the protease by the type II secretion pathway into the extracellular milieu. We also show that the protease undergoes a maturation process with the aid of a secreted factor(s). Finally, we propose that V. cholerae is a collagenovorous bacterium, as it is able to utilize collagen as a sole nutrient source. This study initiates new lines of investigations aiming to uncover the structural and functional components of the V. cholerae collagen utilization program.  相似文献   
2.
Richard A. Gill 《Plant and Soil》2014,374(1-2):197-210

Background and aims

Drivers of ecosystem dynamics that are under human influence range from local, land-management decisions to global processes such as warming temperatures and N deposition. The goal of this study was to understand how multiple, potentially interacting factors influence net primary production, N mineralization, and water and soil CO2 fluxes.

Methods

Here I report on a three-year experiment that manipulated air temperature using ITEX passive warming cones and N deposition in a mountain meadow ecosystems that were historically grazed or protected from grazing.

Results

The strongest and most consistent effect was due to the legacy of grazing, with previously grazed sites having lower primary production, lower soil respiration rates, lower soil moisture, and lower soil C and N stocks than historically ungrazed sites. Warming increased soil respiration, but the effect was transient, and decreased over the 3-year study. Nitrogen addition increased primary production in the second and third year of the experiment but had no significant effect on soil respiration. The effect of historical grazing on primary production was approximately double the effect of N addition. Temperature and N deposition rarely interacted except for increasing N availability during the warm, wet growing season of 2004.

Conclusions

These findings indicate that the legacies of land use, with their influence on plant community composition and hydrologic processes, are locally more important than short-term step changes in temperature and nutrient availability.  相似文献   
3.

Aims

This study investigated Cu uptake and accumulation as well as physiological and biochemical changes in grapevines grown in soils containing excess Cu.

Methods

The grapevines were collected during two productive cycles from three vineyards with increasing concentrations of Cu in the soil and at various growth stages, before and after the application of Cu-based fungicides. The Cu concentrations in the grapevine organs and the macronutrients and biochemical parameters in the leaf blades were analyzed.

Results

At close to the flowering stage of the grapevines, the concentration and content of Cu in the leaves were increased. However, the Cu concentrations in the roots, stem, shoots and bunches did not correlate with the metal concentrations in the soil. The application of Cu-based fungicides to the leaves increased the Cu concentrations in the shoots, leaves and rachis; however, the effect of the fungicides on the Cu concentration in the berries was not significant. The biochemical analyses of the leaf blades demonstrated symptoms of oxidative stress that correlated with the Cu concentrations in soil.

Conclusions

The increased availability of Cu in soil had a slight effect on the levels and accumulation of Cu in mature grapevines during the productive season and did not alter the nutritional status of the plant. However, increased Cu concentrations were observed in the leaves. The evidence of oxidative stress in the leaves correlated with the increased levels of Cu in soil.  相似文献   
4.
5.
The carbon dioxide (CO2)-concentrating mechanism of cyanobacteria is characterized by the occurrence of Rubisco-containing microcompartments called carboxysomes within cells. The encapsulation of Rubisco allows for high-CO2 concentrations at the site of fixation, providing an advantage in low-CO2 environments. Cyanobacteria with Form-IA Rubisco contain α-carboxysomes, and cyanobacteria with Form-IB Rubisco contain β-carboxysomes. The two carboxysome types have arisen through convergent evolution, and α-cyanobacteria and β-cyanobacteria occupy different ecological niches. Here, we present, to our knowledge, the first direct comparison of the carboxysome function from α-cyanobacteria (Cyanobium spp. PCC7001) and β-cyanobacteria (Synechococcus spp. PCC7942) with similar inorganic carbon (Ci; as CO2 and HCO3) transporter systems. Despite evolutionary and structural differences between α-carboxysomes and β-carboxysomes, we found that the two strains are remarkably similar in many physiological parameters, particularly the response of photosynthesis to light and external Ci and their modulation of internal ribulose-1,5-bisphosphate, phosphoglycerate, and Ci pools when grown under comparable conditions. In addition, the different Rubisco forms present in each carboxysome had almost identical kinetic parameters. The conclusions indicate that the possession of different carboxysome types does not significantly influence the physiological function of these species and that similar carboxysome function may be possessed by each carboxysome type. Interestingly, both carboxysome types showed a response to cytosolic Ci, which is of higher affinity than predicted by current models, being saturated by 5 to 15 mm Ci. This finding has bearing on the viability of transplanting functional carboxysomes into the C3 chloroplast.Cyanobacteria inhabit a diverse range of ecological habitats, including both freshwater and marine ecosystems. The flexibility to occupy these different habitats is thought to come in part from the carbon-concentrating mechanism (CCM) present in all species (Badger et al., 2006). The CCM comprises inorganic carbon (Ci; as carbon dioxide [CO2] and HCO3) transporters for Ci uptake and protein microbodies called carboxysomes for CO2 concentration and fixation by Rubisco (Badger and Price, 2003). The CCM is believed to have evolved in response to changes in the absolute and relative levels of CO2 and oxygen (O2) in the atmosphere during the evolution of oxygenic photosynthesis in cyanobacteria (Price et al., 2008).There are two main phylogenetic groups within the cyanobacteria based on Rubisco and carboxysome phylogenies; α-cyanobacteria have α-carboxysomes with Form-IA Rubisco, whereas β-cyanobacteria have β-carboxysomes with Form-IB Rubisco (Tabita, 1999; Badger et al., 2002). Rubisco large subunit protein sequences from these two groups are closely related but nevertheless, distinguishable (Supplemental Fig. S1). In general, α-cyanobacteria and β-cyanobacteria occupy a quite different range of ecological habitats. The α-cyanobacteria are mostly marine organisms, with the majority of species living in the open ocean (Badger et al., 2006). Marine α-cyanobacteria live in very stable environments with high pH (pH 8.2) and dissolved carbon levels but low nutrients. They are characterized by small cells, very small genomes (1.6–2.8 Mb), and a few constitutively expressed carbon uptake transporters (Rae et al., 2011; Beck et al., 2012). They have been described as low flux, low energy cyanobacteria with a minimal CCM (Badger et al., 2006). Although these species are slow growing, oceanic cyanobacteria contribute as much as one-half of oceanic primary productivity (Liu et al., 1997, 1999; Field et al., 1998), suggesting that they may contribute up to 25% to net global productivity every year.In comparison, β-cyanobacteria occupy a much more diverse range of habitats, including freshwater, estuarine, and hot springs and never reach the same levels of global abundance (Badger et al., 2006). They are characterized by larger cells, larger genomes (2.2–3.6 Mb), and an array of carbon uptake transporters, including those transporters induced under low Ci (Rae et al., 2011, 2013). In addition to these broadly defined α-groups and β-groups, there are small numbers of α-cyanobacteria that have been termed transitional strains (Price, 2011; Rae et al., 2011). These species (e.g. Cyanobium spp. PCC7001, Synechococcus spp. WH5701, and Cyanobium spp. PCC6307; Supplemental Fig. S1) live in marginal marine and freshwater environments and have a number of characteristics similar to β-cyanobacteria. For example, they have a more diverse range of Ci uptake systems and a significantly larger genome than closely related α-cyanobacteria, and it has been suggested that the additional genes encoding transport systems were acquired by horizontal gene transfer (HGT) from β-cyanobacteria (Rae et al., 2011).Although the carboxysomes from α-cyanobacteria and β-cyanobacteria are very similar in overall structure, in that they share an outer protein shell of common phylogenetic origin (Kerfeld et al., 2005), they are distinguished from each other largely by differences in the proteins, which seem to make up or interact with the interior of the carboxysome compartment (Supplemental Table S1). This finding suggests that their different structures today have arisen through periods of common and convergent evolution. Certain carboxysome shell proteins from α-carboxysomes and β-carboxysomes show regions of significant sequence homology. These proteins are denoted as CsoS1 to CsoS4 (in α-cyanobacteria) and CcmKLO (in β-cyanobacteria), and the homologous regions have been termed bacterial microcompartment domains (Kerfeld et al., 2010; Rae et al., 2013). Proteins with these domains are also found in bacterial microcompartments in proteobacteria. However, other identified carboxysome proteins do not show any sequence homology between α-carboxysomes and β-carboxysomes but may perform similar functional roles. For example, carbonic anhydrase activity is essential for carboxysome function, but its activity seems to be provided by a range of different proteins (β-CcaA, β-CcmM, and α-CsoSCA; Kupriyanova et al., 2013). Similarly, β-CcmM and α-CsoS2 could play similar roles in organizing the interface between the shell and Rubisco within the carboxysomes (Gonzales et al., 2005; Long et al., 2007).The functioning of a carboxysome relies on a number of biochemical properties associated with the protein microbody structure. These properties include the biochemical/kinetic properties of Rubisco contained within carboxysomes, the conductance of the carboxysome shell to the influx of substrate ribulose-1,5-bisphosphate (RuBP) and the efflux of the carboxylation product phosphoglycerate (PGA), the conductance of the shell to the influx of bicarbonate and the efflux of CO2, and lastly, the manner in which bicarbonate is converted to CO2 within the carboxysomes. α-Carboxysomes and β-carboxysomes have the potential to differ in each of these properties. The flux of phosphorylated sugars across the shell has been postulated to be mediated by the pores in the hexameric shell proteins (Yeates et al., 2010; Kinney et al., 2011), which although similar, do differ between the two carboxysomes types. Bicarbonate and CO2 uptake processes are less well-defined but probably involve aspects of the way in which unique shell interface proteins interact with Rubisco, which also differs in that CsoS2 and CsoSCA are probably the interacting proteins involved in α-carboxysomes (Espie and Kimber, 2011), whereas CcmM and β-carboxysomal CA are variably involved in β-carboxysomes (Long et al., 2010). Finally, the Form-IA and Form-IB Rubisco proteins at the heart of carboxylation, although similar, have the potential to show different kinetic properties. Although Form-IB Rubiscos from β-cyanobacteria are well-characterized, the Form-IA counterparts have received very little attention. In addition, the CCM of very few strains of cyanobacteria have been studied at the level of biochemistry and physiology, and they have been almost exclusively β-cyanobacteria. As a result, there are significant gaps in our knowledge about the similarities and differences in functional traits between α-cyanobacterial and β-cyanobacterial strains. One important question that remains to be answered is whether α-carboxysomes and β-carboxysomes have intrinsic differences in their biochemical properties that influence the nature of the CCM, which is established within each broad cell type.Because of the difficulties in isolating and assaying intact carboxysomes in vitro, the characterization of biochemical properties of carboxysomes is not easily addressed. One way forward is to study the properties of the CCM in detail in a model representative strain from each group and compare their characteristics to contrast the intracellular function of α-cell types and β-cell types. In the past, it has been restricted because of the difficulties in growing many of the open ocean α-cyanobacteria and their very different natures in relation to inorganic transporter composition. However, the availability of α-cyanobacteria transition strains, which grow well in the laboratory, has provided an opportunity to address this question. The α-cyanobacteria Cyanobium spp. PCC7001 (hereafter Cyanobium spp.), in particular, grows in standard freshwater media (BG11) and has growth and photosynthetic performance properties that closely match the model β-cyanobacteria, Synechococcus spp. PCC7942 (hereafter Synechococcus spp.); for this reason, Cyanobium spp. is ideal for a balanced comparison of the in vivo physiological properties of α-carboxysomes and β-carboxysomes in two species with relatively similar Ci-uptake properties.Genome analysis of both strains indicates that Cyanobium spp. have many of the same carbon uptake systems present in Synechococcus spp. (Rae et al., 2011). In using two strains with such similar transport capacities, we aimed to shed light on aspects of the functional properties of carboxysomes in each strain and how these properties affect the operation of the CCM in α-cyanobacteria and β-cyanobacteria. Using both membrane inlet mass spectrometry (MIMS) and silicon oil centrifugation, we investigated Ci pool sizes and CO2 uptake rates in both species for cells grown at high and low CO2. Comparative Rubisco properties and photosynthetic rates of each species were determined, and intracellular pools of RuBP and PGA were measured. In addition, we characterized a number of cellular properties to determine differences in the biochemical environments in which each carboxysome type exists. Together, the results provide a unique functional comparison of two distinct carboxysome types from phylogenetically disparate cyanobacteria.  相似文献   
6.
Opium poppy (Papaver somniferum) is one of the world’s oldest medicinal plants and remains the only commercial source for the narcotic analgesics morphine, codeine and semi-synthetic derivatives such as oxycodone and naltrexone. The plant also produces several other benzylisoquinoline alkaloids with potent pharmacological properties including the vasodilator papaverine, the cough suppressant and potential anticancer drug noscapine and the antimicrobial agent sanguinarine. Opium poppy has served as a model system to investigate the biosynthesis of benzylisoquinoline alkaloids in plants. The application of biochemical and functional genomics has resulted in a recent surge in the discovery of biosynthetic genes involved in the formation of major benzylisoquinoline alkaloids in opium poppy. The availability of extensive biochemical genetic tools and information pertaining to benzylisoquinoline alkaloid metabolism is facilitating the study of a wide range of phenomena including the structural biology of novel catalysts, the genomic organization of biosynthetic genes, the cellular and sub-cellular localization of biosynthetic enzymes and a variety of biotechnological applications. In this review, we highlight recent developments and summarize the frontiers of knowledge regarding the biochemistry, cellular biology and biotechnology of benzylisoquinoline alkaloid biosynthesis in opium poppy.  相似文献   
7.
New data have been acquired on the biology, morphological features and distribution of Norwegian (Atlantic) pollock Theragra finnmarchica in the Barents Sea. Two individuals of this rare species gadoid (Gadidae) were caught in June and July 2012 in the south-eastern part of the Barents Sea, indicating a wider distribution area of this species than previously thought. It has been confirmed that a number of morphological features of Norwegian pollock is different from T. chalcogramma, and that it feeds on macroplankton (Euphausiidae, Hyperiidae).  相似文献   
8.
Characteristics of morphology and number of melanomacrophage centers (MMCs) in the liver and spleen of the roach Rutilus rutilus and the amount of pigments in MMCs during the Haff disease outbreak and the death of fish in Lake Kotokel in relation to these parameters in the roach from Lake Baikal are described. Pathological changes in the microvasculature and parenchyma in the liver of the roach from Lake Kotokel were found. The area of melanomacrophage centers in the liver of the roach from this lake was significantly smaller, whereas the number and size of these centers in the spleen was significantly larger than in the roaches from Lake Baikal. Among the pigments studied, the strongest response to the content of this toxin in the water body was shown by hemosiderin. An increase in its amount in the spleen MMCs testifies to an enhanced degradation of erythrocytes and iron release, which may be caused by the damage of cells of the erythrocyte lineage by the toxin.  相似文献   
9.
Eukaryotic ribosome biogenesis involves ∼200 assembly factors, but how these contribute to ribosome maturation is poorly understood. Here, we identify a network of factors on the nascent 60S subunit that actively remodels preribosome structure. At its hub is Rsa4, a direct substrate of the force-generating ATPase Rea1. We show that Rsa4 is connected to the central protuberance by binding to Rpl5 and to ribosomal RNA (rRNA) helix 89 of the nascent peptidyl transferase center (PTC) through Nsa2. Importantly, Nsa2 binds to helix 89 before relocation of helix 89 to the PTC. Structure-based mutations of these factors reveal the functional importance of their interactions for ribosome assembly. Thus, Rsa4 is held tightly in the preribosome and can serve as a “distribution box,” transmitting remodeling energy from Rea1 into the developing ribosome. We suggest that a relay-like factor network coupled to a mechano-enzyme is strategically positioned to relocate rRNA elements during ribosome maturation.  相似文献   
10.
Cell migration is mediated by the dynamic remodeling of focal adhesions (FAs). Recently, an important role of endosomal signaling in regulation of cell migration was recognized. Here, we show an essential function for late endosomes carrying the p14–MP1 (LAMTOR2/3) complex in FA dynamics. p14–MP1-positive endosomes move to the cell periphery along microtubules (MTs) in a kinesin1- and Arl8b-dependent manner. There they specifically target FAs to regulate FA turnover, which is required for cell migration. Using genetically modified fibroblasts from p14-deficient mice and Arl8b-depleted cells, we demonstrate that MT plus end–directed traffic of p14–MP1-positive endosomes triggered IQGAP1 disassociation from FAs. The release of IQGAP was required for FA dynamics. Taken together, our results suggest that late endosomes contribute to the regulation of cell migration by transporting the p14–MP1 scaffold complex to the vicinity of FAs.  相似文献   
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