Phosphorylation is a major constituent of the CTD code, which describes the set of post-translational modifications on 52 repeats of a YSPTSPS consensus heptad that orchestrates the binding of ...regulatory proteins to the C-terminal domain (CTD) of RNA polymerase II. Phospho-specific antibodies are used to detect CTD phosphorylation patterns. However, their recognition repertoire is underexplored due to limitations in the synthesis of long multiphosphorylated peptides. Herein, we describe the development of a synthesis strategy that provides access to multiphosphorylated CTD peptides in high purity without HPLC purification for immobilization onto microtiter plates. Native chemical ligation was used to assemble 12 heptad repeats in various phosphoforms. The synthesis of >60 CTD peptides, 48–90 amino acids in length and containing up to 6 phosphosites, enabled a detailed and rapid analysis of the binding characteristics of different anti-pSer2 antibodies. The three antibodies tested showed positional selectivity with marked differences in the affinity of the antibodies for pSer2-containing peptides. Furthermore, the length of the phosphopeptides allowed a systematic analysis of the multivalent chelate-type interactions. The absence of multivalency-induced binding enhancements is probably due to the high flexibility of the CTD scaffold. The effect of clustered phosphorylation proved to be more complex. Recognition of pSer2 by anti-pSer2-antibodies can be prevented and, perhaps surprisingly, enhanced by the phosphorylation of “bystander” amino acids in the vicinity. The results have relevance for functional analysis of the CTD in cell biological experiments.
Oil is a major pollutant of the environment, and terrestrial oil spills frequently occur in desert areas. Although arthropods account for a large share of animal diversity, the effect of oil ...pollution on this group is rarely documented. We evaluated the effects of oil pollution on parasitoid wasps associated with Vachellia (formerly Acacia) tortilis (Forssk.) and Vachellia raddiana (Savi) trees in a hyper-arid desert that was affected by two major oil spills (in 1975 and 2014). We sampled the parasitoid populations between 2016 and 2018 in three sampling sites and compared their abundance, diversity, and community composition between oil-polluted and unpolluted trees. Parasitoid abundance in oil-polluted trees was lower in one of the sites affected by the recent oil spill, but not in the site affected by the 1975 oil spill. Oil-polluted trees supported lower parasitoid diversity than unpolluted trees in some sampling site/year combinations; however, such negative effects were inconsistent and pollution explained a small proportion of the variation in parasitoid community composition. Our results indicate that oil pollution may negatively affect parasitoid abundances and diversity, although the magnitude of the effect depends on the tree species, sampling site, and the time since the oil spill.
We present few-femtosecond shadowgraphic snapshots taken during the nonlinear evolution of the plasma wave in a laser wakefield accelerator with transverse synchronized few-cycle probe pulses. These ...snapshots can be directly associated with the electron density distribution within the plasma wave and give quantitative information about its size and shape. Our results show that self-injection of electrons into the first plasma-wave period is induced by a lengthening of the first plasma period. Three-dimensional particle-in-cell simulations support our observations.
All life on earth depends on the generation and exploitation of ionic and pH gradients across membranes. One theory for the origin of life proposes that geological pH gradients were the prebiotic ...ancestors of these cellular disequilibria. With an alkaline interior and acidic exterior, alkaline vents match the topology of modern cells, but it remains unknown whether the steep pH gradients persist at the microscopic scale. Herein, we demonstrate the existence of 6 pH‐unit gradients across micrometer scales in a microfluidic vent replicate. Precipitation of metal sulfides at the interface strengthens the gradients, but even in the absence of precipitates laminar flow sustains the disequilibria. The gradients drive directed transport at the fluid interface, leading to colloid accumulation or depletion. Our results confirm that alkaline vents can provide an exploitable pH gradient, supporting their potential role at the emergence of chemiosmosis and the origin of life.
At the interface: A 6 pH‐unit gradient is shown to persist on the microscale in alkaline hydrothermal vents. Observation and modeling of directed transport by diffusiophoresis in the same setting further supports the potential role of electrochemical gradients in alkaline vents at the emergence of chemiosmosis and the origin of life.
Tea is the world's most popular non-alcoholic beverage. China and India are known to be the largest tea producing countries and recognized as the centers for the domestication of the tea plant ...(Camellia sinensis (L.) O. Kuntze). However, molecular studies on the origin, domestication and relationships of the main teas, China type, Assam type and Cambod type are lacking.
Twenty-three nuclear microsatellite markers were used to investigate the genetic diversity, relatedness, and domestication history of cultivated tea in both China and India. Based on a total of 392 samples, high levels of genetic diversity were observed for all tea types in both countries. The cultivars clustered into three distinct genetic groups (i.e. China tea, Chinese Assam tea and Indian Assam tea) based on STRUCTURE, PCoA and UPGMA analyses with significant pairwise genetic differentiation, corresponding well with their geographical distribution. A high proportion (30%) of the studied tea samples were shown to possess genetic admixtures of different tea types suggesting a hybrid origin for these samples, including the Cambod type.
We demonstrate that Chinese Assam tea is a distinct genetic lineage from Indian Assam tea, and that China tea sampled from India was likely introduced from China directly. Our results further indicate that China type tea, Chinese Assam type tea and Indian Assam type tea are likely the result of three independent domestication events from three separate regions across China and India. Our findings have important implications for the conservation of genetic stocks, as well as future breeding programs.
Celotno besedilo
Dostopno za:
DOBA, IZUM, KILJ, NUK, PILJ, PNG, SAZU, SIK, UILJ, UKNU, UL, UM, UPUK
Owing to their ubiquitous distribution, expected beneficial effects and suspected adverse effects, nanoparticles are viewed as a double-edged sword, necessitating a better understanding of their ...interactions with tissues and organisms. Thus, the goals of the present study were to develop and present a method to generate quantitative data on nanoparticle entry into cells in culture and to exemplarily demonstrate the usefulness of this approach by analyzing the impact of size, charge and various proteinaceous coatings on particle internalization. N9 microglial cells and both undifferentiated and differentiated SH-SY5Y neuroblastoma cells were exposed to customized gold nanoparticles. After silver enhancement, the particles were visualized by epipolarization microscopy and analysed by high-content analysis. The value of this approach was substantiated by assessing the impact of various parameters on nanoparticle uptake. Uptake was higher in microglial cells than in neuronal cells. Only microglial cells showed a distinct size preference, preferring particles with a diameter of 80 nm. Positive surface charge had the greatest impact on particle uptake. Coating with bovine serum albumin, fetuin or protein G significantly increased particle internalization in microglial cells but not in neuronal cells. Coating with wheat germ agglutinin increased particle uptake in both N9 and differentiated SH-SY5Y cells but not in undifferentiated SH-SY5Y cells. Furthermore, internalization was shown to be an active process and indicators of caspase-dependent apoptosis revealed that gold nanoparticles did not have any cytotoxic effects. The present study thus demonstrates the suitability of gold nanoparticles and high-content analysis for assessing numerous variables in a stringently quantitative and statistically significant manner. Furthermore, the results presented herein showcase the feasibility of specifically targeting nanoparticles to distinct cell types.
High-content analysis is a powerful tool to quantitate the impact of various factors affecting the uptake of untagged gold nanoparticles.
Acacia trees are keystone species in many arid environments, supporting high levels of plant and animal diversity. In Israel, the populations of Vachellia (formerly Acacia) tortilis (Forssk.) and V. ...raddiana (Savi) are declining at an alarming rate. Severe infestations by bruchid beetles (Coleoptera, Chrysomelidae) are among the major causes of seed mortality, but additional environmental stressors can reduce the defence level of the seeds, exacerbating their susceptibility to predators. In a hyper-arid desert ecosystem affected by two major oil spills (in 1975 and 2014), we quantified seed predation rates caused by insect granivores before and after the pods dropped to the ground. We recorded predation rates of up to 84% for both tree species, and higher predation rates at the ground level than in the canopy, suggesting that repeated infestations occur. These results reinforce the call to protect the populations of large ungulates such as gazelles, which kill the bruchids by feeding upon the pods, and promote seed germination and dispersion. We found no clear evidence of a negative effect of the oil spill on seed predation, indicating that oil pollution did not increase the vulnerability of the seeds to granivores even in trees affected by the recent oil spill.
Celotno besedilo
Dostopno za:
DOBA, IZUM, KILJ, NUK, PILJ, PNG, SAZU, UILJ, UKNU, UL, UM, UPUK
The weakly coordinated triflate complex (P∧P)Pd(OTf)+(OTf)− (1) (P∧P = 1,3-bis(di-tert-butylphosphino)propane) is a suitable reactive precursor for mechanistic studies of the isomerizing ...alkoxcarbonylation of methyl oleate. Addition of CH3OH or CD3OD to 1 forms the hydride species (P∧P)PdH(CH3OH)+(OTf)− (2-CH 3 OH) or the deuteride (P∧P)PdD(CD3OD)+(OTf)− (2 D -CD 3 OD), respectively. Further reaction with pyridine cleanly affords the stable and isolable hydride (P∧P)PdH(pyridine)+(OTf)− (2-pyr). This complex yields the hydride fragment free of methanol by abstraction of pyridine with BF3·OEt2, and thus provides an entry to mechanistic observations including intermediates reactive toward methanol. Exposure of methyl oleate (100 equiv) to 2 D -CD 3 OD resulted in rapid isomerization to the thermodynamic isomer distribution, 94.3% of internal olefins, 5.5% of α,β-unsaturated ester and <0.2% of terminal olefin. Reaction of 2-pyr/BF3·OEt2 with a stoichiometric amount of 1-13C-labeled 1-octene at −80 °C yields a 50:50 mixture of the linear alkyls (P∧P)Pd13 CH2(CH2)6CH3+ and (P∧P)PdCH2(CH2)6 13 CH3+ (4a and 4b). Further reaction with 13CO yields the linear acyls (P∧P)Pd13C(O)12/13CH2(CH2)6 12/13CH3(L)+ (5-L; L = solvent or 13CO). Reaction of 2-pyr/BF3·OEt2 with a stoichiometric amount of methyl oleate at −80 °C also resulted in fast isomerization to form a linear alkyl species (P∧P)PdCH2(CH2)16C(O)OCH3+ (6) and a branched alkyl stabilized by coordination of the ester carbonyl group as a four membered chelate (P∧P)PdCH{(CH2)15CH3}C(O)OCH3+ (7). Addition of carbon monoxide (2.5 equiv) at −80 °C resulted in insertion to form the linear acyl carbonyl (P∧P)PdC(O)(CH2)17C(O)OCH3(CO)+ (8-CO) and the five-membered chelate (P∧P)PdC(O)CH{(CH2)15CH3}C(O)OCH3+ (9). Exposure of 8-CO and 9 to 13CO at −50 °C results in gradual incorporation of the 13C label. Reversibility of 7 + CO ⇄ 9 is also evidenced by ΔG = −2.9 kcal mol–1 and ΔG ⧧ = 12.5 kcal mol–1 from DFT studies. Addition of methanol at −80 °C results in methanolysis of 8-L (L = solvent) to form the linear diester, 1,19-dimethylnonadecandioate, whereas 9 does not react and no branched diester is observed. DFT yields a barrier for methanolysis of ΔG ⧧ = 29.7 kcal mol–1 for the linear (8) vs ΔG ⧧ = 37.7 kcal mol–1 for the branched species (9).
We present a divergent strategy for the fluorination of phenylacetic acid derivatives that is induced by a charge-transfer complex between Selectfluor and 4-(dimethylamino)pyridine. A comprehensive ...investigation of the conditions revealed a critical role of the solvent on the reaction outcome. In the presence of water, decarboxylative fluorination through a single-electron oxidation is dominant. Non-aqueous conditions result in the clean formation of α-fluoro-α-arylcarboxylic acids.
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