Proteomic characterization of hierarchical megacomplex formation in Arabidopsis thylakoid membrane

Conversion of solar energy into chemical energy in plant chloroplasts concomitantly modifies the thylakoid architecture and hierarchical interactions between pigment–protein complexes. Here, the thylakoids were isolated from light‐acclimated Arabidopsis leaves and investigated with respect to the composition of the thylakoid protein complexes and their association into higher molecular mass complexes, the largest one comprising both photosystems (PSII and PSI) and light‐harvesting chlorophyll a/b‐binding complexes (LHCII). Because the majority of plant light‐harvesting capacity is accommodated in LHCII complexes, their structural interaction with photosystem core complexes is extremely important for efficient light harvesting. Specific differences in the strength of LHCII binding to PSII core complexes and the formation of PSII supercomplexes are well characterized. Yet, the role of loosely bound L‐LHCII that disconnects to a large extent during the isolation of thylakoid protein complexes remains elusive. Because L‐LHCII apparently has a flexible role in light harvesting and energy dissipation, depending on environmental conditions, its close interaction with photosystems is a prerequisite for successful light harvesting in vivo. Here, to reveal the labile and fragile light‐dependent protein interactions in the thylakoid network, isolated membranes were subjected to sequential solubilization using detergents with differential solubilization capacity and applying strict quality control. Optimized 3D‐lpBN‐lpBN‐sodium dodecyl sulfate–polyacrylamide gel electrophoresis system demonstrated that PSII–LHCII supercomplexes, together with PSI complexes, hierarchically form larger megacomplexes via interactions with L‐LHCII trimers. The polypeptide composition of LHCII trimers and the phosphorylation of Lhcb1 and Lhcb2 were examined to determine the light‐dependent supramolecular organization of the photosystems into megacomplexes.     

Transient trade‐off between climate benefit and biodiversity loss of harvesting stumps for bioenergy

To replace fossil fuel and thereby mitigate climate change, harvesting of wood such as stumps for bioenergy will likely increase. Coarse deadwood is an important resource for biodiversity and stumps comprise the main part of the coarse deadwood in managed forests. We provide the first integrated analysis of the long‐term climate and biodiversity impacts of a whole landscape. We simultaneously project climate and biodiversity impacts of harvesting stumps to substitute for fossil coal, assuming scenarios with different proportions of the landscape with stump harvest (10, 50, 80%) the coming 50 years. A life cycle approach was used to calculate future global temperature changes and future metapopulation changes in six epixylic lichens. Metapopulation dynamics were projected using colonization and extinction models based on times series data. Harvesting stumps from ≥50% of the clear‐cut forest land benefits climate with a net global temperature reduction >0.5·10^?9 K ha^?1 after 50 years if assuming substitution of fossil coal. For all scenarios, using stump bioenergy leads to immediate (within 1 year) reductions in temperature of 50% compared to using fossil coal, increasing to 70% reduction after 50 years. However, large‐scale stump harvest inflicted substantial metapopulation declines for five of six lichens. High stump harvest levels (≥50%) put common lichens at risk of becoming red‐listed following the IUCN criteria. The net temperature reduction (cooling effect) from substituting fossil coal with stumps harvested for bioenergy increased over time, while lichen metapopulations stabilized at lower equilibria after two to three decades. This indicates that trade‐offs between climate and metapopulations of commons species are transient, where climate benefits become more prevalent in the long term. As both objectives are important for meeting (inter‐)national climate and biodiversity targets, integrated analyses such as this should be encouraged and urged to guide policymaking about large‐scale implementation of stump harvest.     

The Effect of the −308A Allele of the TNF‐α Gene on Insulin Action Is Dependent on Obesity

Objective: Promoter polymorphisms of the tumor necrosis factor alpha (TNF‐α) gene are associated with insulin sensitivity and BMI. We investigated whether the effect of the G‐308A polymorphism of the TNF‐α gene on insulin action depends on BMI. Research Methods and Procedures: The effects of the G‐308A polymorphism on the rates of glucose and lipid oxidation and free fatty acid (FFA) levels were studied using the hyperinsulinemic euglycemic clamp combined with indirect calorimetry in 129 healthy subjects. Results: The ?308A allele of the TNF‐α gene was associated with high rates of glucose oxidation (p = 0.008 adjusted for age, gender, and BMI) and lipid synthesis (p = 0.037) and suppression of FFA levels (p = 0.023) during hyperinsulinemia. In normal weight subjects (BMI < 26 kg/m²), the ?308 allele was associated with high rates of glucose oxidation (p = 0.036) during the clamp but not with high rates of lipid synthesis (p = 0.896) or FFA suppression (p = 0.464). In overweight subjects (BMI ≥ 26 kg/m²), high rates of lipid synthesis and FFA suppression (p = 0.010 and p = 0.042, respectively) but not the rates of glucose oxidation during the clamp (p = 0.193) were associated with the ?308A allele. Discussion: The ?308A allele of the promoter of the TNF‐α gene is associated with high rates of glucose oxidation in normal weight subjects and with effective lipid storage in overweight subjects. These findings suggest an interaction of the polymorphism with obesity.     

Differential expression of granulysin and perforin by NK cells in cancer patients and correlation of impaired granulysin expression with progression of cancer

Granulysin has been identified as an effector molecule co-localized with perforin in the cytotoxic granules of cytotoxic T lymphocytes and natural killer (NK) cells, and has been reported to kill intracellular pathogens in infected cells in the presence of perforin and to induce a cytotoxic effect against tumor cells. The aim of the present study was to elucidate whether intracellular expression of granulysin and perforin by NK cells might be associated with progression of cancer. Flow cytometric analysis demonstrated high levels of perforin and granulysin expression by CD3(-) CD16(+) cells in healthy controls. In contrast, cancer patients exhibited significantly decreased levels of granulysin expression ( P<0.005), despite having equally high levels of perforin expression in comparison with healthy controls. The tumor-free patients expressed granulysin at levels similar to healthy controls, while the progressive tumor-bearing patients expressed remarkably lower levels of granulysin compared to healthy controls ( P<0.0001). Similarly, patients with an advanced performance status had significantly fewer granulysin-positive NK cells than healthy controls. Meanwhile, a considerable number of the tumor-bearing patients showed a decrease in the number of circulating NK cells, and a correlation between impaired granulysin expression and reduced circulating NK cells was observed. These findings suggest that the tumor-bearing patients with impaired granulysin expression were in an immunosuppressive state. In conclusion, impaired expression of granulysin by NK cells correlates with progression of cancer, and determination of granulysin expression might prove informative for assessing the immunological condition of cancer patients.     

The trans Golgi Network Is Lost from Cells Infected with African Swine Fever Virus

The cellular secretory pathway is important during the assembly and envelopment of viruses and also controls the transport of host proteins, such as cytokines and major histocompatibility proteins, that function during the elimination of viruses by the immune system. African swine fever virus (ASFV) encodes at least 26 proteins with stretches of hydrophobic amino acids suggesting entry into the secretory pathway (R. J. Yanez, J. M. Rodriguez, M. L. Nogal, L. Yuste, C. Enriquez, J. F. Rodriguez, and E. Vinuela, Virology 208:249-278, 1995). To predict how and where these potential membrane proteins function, we have studied the integrity of the secretory pathway in cells infected with ASFV. Remarkably, ASFV caused complete loss of immunofluorescence signal for the trans Golgi network (TGN) marker protein TGN46 and dispersed the AP1 TGN adapter complex. Loss of TGN46 signal was not due to degradation of TGN46, suggesting redistribution of TGN46 to other membrane compartments. ASFV markedly slowed transport of cathepsin D to lysosomes, demonstrating that loss of TGN structure correlated with loss of TGN function. ASFV shows a tropism for macrophages, and it is possible that ASFV compromises TGN function to augment the activity of viral membrane proteins or to suppress the function of host immunoregulatory proteins.     

Establishment of a Seronegative Human T-Cell Leukemia Virus Type 1 (HTLV-1) Carrier State in Rats Inoculated with a Syngeneic HTLV-1-Immortalized T-Cell Line Preferentially Expressing Tax

Human T-cell leukemia virus type 1 (HTLV-1) causes T-cell malignancies in a small percentage of the population infected with the virus after a long carrier state. In the present study, we established a seronegative HTLV-1 carrier state in rats inoculated with a newly established HTLV-1-infected rat T cell line, FPM1. FPM1 originated from rat thymocytes cocultured with a human HTLV-1 producer, MT-2 cells, and expressed rat CD4, CD5, CD25, and HTLV-1 Tax. However, FPM1 scarcely expressed other major HTLV-1 structural proteins and failed to induce typical antibody responses against HTLV-1 in inoculated rats. In contrast, control rats inoculated with MT-2 cells generated significant levels of anti-HTLV-1 antibodies. HTLV-1 proviruses were detected in peripheral blood cells of syngeneic rats inoculated with FPM1 for more than 1 year. Analysis of the flanking region of HTLV-1 provirus integrated into host cells suggested that FPM1 cells remained in these animals over a relatively long period of time. However, a similar seronegative HTLV-1 carrier state was induced in the rats inoculated with mitomycin C-treated FPM1 cells and also in FPM1-inoculated allogeneic rats, suggesting that FPM1 could also transmit HTLV-1 into host cells in vivo. Our findings indicated that (i) HTLV-1-immortalized T cells which preferentially express HTLV-1 Tax persisted in vivo but failed to induce any diseases in immunocompetent syngeneic rats and that (ii) suboptimal levels of HTLV-1 for antibody responses allowed the establishment of persistent HTLV-1 infection.     

Homoplasy Increases Phylogenetic Structure

According to currently accepted theories, rapidly evolving nucleotide sites are phylogenetically less informative than more slowly evolving ones, especially for recognizing more ancient groupings. For this reason third codon positions are often regarded as less reliable than first and second positions as indicators of phylogeny. Analysis of the largest nucleotide matrix treated to date—2538 rbc L sequences covering all major lineages of green plants—shows the opposite: although rapidly evolving and highly homoplastic, third positions contain most of the phylogenetic structure in the data. Frequency of change should thus be used with caution as a criterion for weighting or selecting characters.     

Sexual dimorphism, sex ratio and spatial distribution of male and female shrubs in the dioecious speciesPistacia lentiscus L.

The population sex ratio, spatial distribution, relative growth rate, canopy size and morphology, and nitrogen content of leaves of male and female individuals were compared in seven populations of the dioecious shrubPistacia lentiscus L. occurring in different localities of the Serra da Arrábida Natural Park, Portugal. The expected pattern of male preponderance in stressful habitats was tested. Less perturbed areas, with a well developed vegetation cover, had male-biased sex ratios, while in former agricultural areas there were no significant differences between the number of male and female plants. Our results failed to reveal any significant differences in the morphological variables between sexes except in the number of basal stems. Male plants also had a significantly higher chlorophyll content (chl a+b) than female plants. But when plants were sorted into large and small individuals, larger plants showed a significantly higher leaf area index and a significantly higher number of basal stems. There were no significant differences in the leaf nitrogen content between male and female plants from June to August, but such differences appeared (P<0.001) in September and October corresponding to the production of mature fruits. Differences in sex-ratio may be explained by different resource allocation patterns of the two sexes, which favoured male plants in the less perturbed environments. In agricultural areas farmers could also have favoured female plants in former times.