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The Rab Geranylgeranyl Transferase Beta Subunit Is Essential for Embryo and Seed Development in Arabidopsis thaliana
Auxin is a key regulator of plant development affecting the formation and maturation of reproductive structures. The apoplastic route of auxin transport engages influx and efflux facilitators from the PIN, AUX and ABCB families. The polar localization of these proteins and constant recycling
from the plasma membrane to endosomes is dependent on Rab-mediated vesicular traffic. Rab proteins are anchored to membranes via posttranslational addition of two geranylgeranyl moieties by the Rab Geranylgeranyl Transferase enzyme (RGT), which consists of RGTA, RGTB and REP subunits. Here, we present data showing that seed development in the rgtb1 mutant, with decreased vesicular transport capacity, is disturbed. Both pre- and post-fertilization events are affected, leading to a decrease in seed yield. Pollen tube recognition at the stigma and its guidance to the micropyle is compromised and the seed coat forms incorrectly. Excess auxin in the sporophytic tissues of the ovule in the rgtb1 plants leads to an increased tendency of autonomous endosperm formation in unfertilized
ovules and influences embryo development in a maternal sporophytic manner. The results show the importance of vesicular traffic for sexual reproduction in flowering plants, and highlight RGTB1 as a key component of sporophytic-filial signaling
The GTPase Arf1 Is a Determinant of Yeast Vps13 Localization to the Golgi Apparatus
VPS13 proteins are evolutionarily conserved. Mutations in the four human genes (VPS13A-D) encoding VPS13A-D proteins are linked to developmental or neurodegenerative diseases. The relationship between the specific localization of individual VPS13 proteins, their molecular functions, and the pathology of these diseases is unknown. Here we used a yeast model to establish the determinants of Vps13′s interaction with the membranes of Golgi apparatus. We analyzed the different phenotypes of the arf1-3 arf2Δ vps13∆ strain, with reduced activity of the Arf1 GTPase, the master regulator of Golgi function and entirely devoid of Vps13. Our analysis led us to propose that Vps13 and Arf1 proteins cooperate at the Golgi apparatus. We showed that Vps13 binds to the Arf1 GTPase through its C-terminal Pleckstrin homology (PH)-like domain. This domain also interacts with phosphoinositol 4,5-bisphosphate as it was bound to liposomes enriched with this lipid. The homologous domain of VPS13A exhibited the same behavior. Furthermore, a fusion of the PH-like domain of Vps13 to green fluorescent protein was localized to Golgi structures in an Arf1-dependent manner. These results suggest that the PH-like domains and Arf1 are determinants of the localization of VPS13 proteins to the Golgi apparatus in yeast and humans
Effect of salt stress in urban conditions on two Acer species with different sensitivity
Background. The benefits of trees in urban areas include the following: an increase in ecosystem health, an increase in human health, the mitigation of the effects of heat and drought at microclimate level, the storage and sequestration of carbon, and a reduction in air pollution and noise. These ecosystem services can be provided only by trees that are in good health. The main cause of salt stress in urban environments is the use of deicing salts on the streets in winter. Salt stress is a complex process that includes changes in plants on the physiological, histological, cellular and molecular levels, leading to limitations in nutrient uptake, disrupting the ionic balance of trees and resulting in the death of roadside trees. In response to salinity, trees have developed a variety of defence mechanisms that allow them to minimize the effects of stress and maintain homeostasis.
Methodology. The reactions of two species Acer species: A. platanoides and A. campestre, which have different sensitivities to the unfavourable conditions of the
urban environments (mainly salt stress), were investigated. The research included two experiments: a field experiment with city trees and a controlled pot experiment with young trees treated with increasing doses of salt. In both experiments, the following were performed: an assessment of the health condition of the trees and the content of macroelements as well as the Cl and Na in leaves and a qualitative and quantitative analysis of polyprenols.
Results. A. campestre had a more specific strategy than A. platanoides for dealing with Na and Cl, which resulted in undamaged leaves. Under the same conditions, A. platanoides leaves contained more Cl and Na and were severely damaged. The disruption of the ion balance due to salt stress was lower in A. campestre than in A. platanoides. Compared with A. platanoides, A. campestre synthesized more polyprenols in the field experiment. This ability was acquired during the process of acclimation, because it occurred only in the mature trees in the field experiment and not in the young trees in the pot experiment.
Conclusions. The use of two experimental methods (i.e., the field and pot experiments) allowed for a more complete assessment of tree strategies to mitigate salt stress. A. campestre displayed a more specific strategy than A. platanoides. This strategy was based on several elements. A. campestre limited Cl and Na transport to the leaves, which resulted in a lack of damage to those organs. Under the same conditions, A. platanoides individuals contained more Cl and Na in their leaves and were seriously damaged. A. campestre synthesized larger amounts of polyprenols, which probably have the ability to mitigate salt stress. This ability was acquired during the process of acclimation, because it occurred only in the mature trees in the field experiment and was not observed in the young trees in the pot experiment
Control of ABA Signaling and Crosstalk with Other Hormones by the Selective Degradation of Pathway Components
A rapid and appropriate genetic and metabolic acclimation, which is crucial for plants’survival in a changing environment, is maintained due to the coordinated action of plant hormones and cellular degradation mechanisms influencing proteostasis. The plant hormone abscisic acid
(ABA) rapidly accumulates in plants in response to environmental stress and plays a pivotal role in the reaction to various stimuli. Increasing evidence demonstrates a significant role of autophagy in controlling ABA signaling. This field has been extensively investigated and new discoveries are constantly being provided. We present updated information on the components of the ABA signaling pathway, particularly on transcription factors modified by different E3 ligases. Then, we focus on the role of selective autophagy in ABA pathway control and review novel evidence on the involvement of autophagy in different parts of the ABA signaling pathway that are important for crosstalk with other hormones, particularly cytokinins and brassinosteroids
Phosphorylation impacts Cu(II) binding by ATCUN motifs
ATCUN (amino terminal Cu(II) and Ni(II) binding) motifs chelate Cu(II) ions strongly. However, the impact of the phosphorylation of neighboring residues on such complexation has not been elucidated. The copper(II) dissociation constants of original and phosphorylated peptides from human histatin-1 and human serum albumin were compared using spectroscopic methods. Phosphorylation markedly weakened Cu(II) binding. Thus, these results indicate that phosphorylation may be a vital mechanism governing metal ion binding
Proteome Analysis of PC12 Cells Reveals Alterations in Translation Regulation and Actin Signaling Induced by Clozapine
Improved HDAC Inhibition, Stronger Cytotoxic Effect and Higher Selectivity against Leukemias and Lymphomas of Novel, Tricyclic Vorinostat Analogues
Histone deacetylase (HDAC) inhibitors are a class of drugs used in the cancer treatment. Here, we developed a library of 19 analogues of Vorinostat, an HDAC inhibitor used in lymphomas treatment. In Vorinostat, we replaced the hydrophobic phenyl group with various tricyclic ‘caps’ possessing a central, eight-membered, heterocyclic ring, and investigated the HDAC activity and cytotoxic effect on the cancer and normal cell lines. We found that 3 out of the 19 compounds, based on dibenzo[b,f]azocin-6(5H)-one, 11,12-dihydrodibenzo[b,f]azocin- 6(5H)-one, and benzo[b]naphtho[2,3-f][1,5]diazocine-6,14(5H,13H)-dione scaffolds, showed better HDACs inhibition than the referenced Vorinostat. In leukemic cell line MV4-11 and in the lymphoma cell line Daudi, three compounds showed lower IC50 values than Vorinostat. These compounds had higher activity and selectivity against MV4-11 and Daudi cell lines than reference Vorinostat. We also observed a strong correlation between HDACs inhibition and the cytotoxic effect. Cell lines derived from solid tumours: A549 (lung carcinoma) and MCF-7 (breast adenocarcinoma) as well as reference BALB/3T3 (normal murine fibroblasts) were less susceptible to compounds tested. Developed derivatives show improved properties than Vorinostat, thus they could be considered as possible agents for leukemia and lymphoma treatment
Pharmacokinetic Drug Interaction Study of Sorafenib and Morphine in Rats
A combination of the tyrosine kinase inhibitor—sorafenib—and the opioid analgesic—
morphine—can be found in the treatment of cancer patients. Since both are substrates of Pglycoprotein
(P-gp), and sorafenib is also an inhibitor of P-gp, their co-administration may affect their
pharmacokinetics, and thus the safety and efficacy of cancer therapy. Therefore, the aim of this study
was to evaluate the potential pharmacokinetic drug–drug interactions between sorafenib and morphine
using an animal model. The rats were divided into three groups that Received: sorafenib and
morphine (ISOR+MF), sorafenib (IISOR), and morphine (IIIMF). Morphine caused a significant increase
in maximum plasma concentrations (Cmax) and the area under the plasma concentration–time curves
(AUC0–t, and AUC0–¥) of sorafenib by 108.3 (p = 0.003), 55.9 (p = 0.0115), and 62.7% (p = 0.0115),
respectively. Also, the Cmax and AUC0–t of its active metabolite—sorafenib N-oxide—was significantly
increased in the presence of morphine (p = 0.0022 and p = 0.0268, respectively). Sorafenib, in
turn, caused a significant increase in the Cmax of morphine (by 0.5-fold, p = 0.0018). Moreover, in the
presence of sorafenib the Cmax, AUC0–t, and AUC0–¥ of the morphine metabolite M3G increased
by 112.62 (p < 0.0001), 46.82 (p = 0.0124), and 46.78% (p = 0.0121), respectively. Observed changes
in sorafenib and morphine may be of clinical significance. The increased exposure to both drugs
may improve the response to therapy in cancer patients, but on the other hand, increase the risk of
adverse effects
Patterns of Diversity of Fusarium Fungi Contaminating Soybean Grains
Soybean is an important, high protein source of food and feed. However, like other agricultural grains, soybean may pose a risk to human and animal health due to contamination of the grains with toxigenic Fusaria and associated mycotoxins. In this study, we investigated the diversity of Fusaria on a panel of 104 field isolates obtained from soybean grains during the growing seasons in 2017–2020. The results of species-specific PCR analyses showed that Fusarium avenaceum was the most common (n = 40) species associated with soybean grains in Poland, followed by F. equiseti (n = 22) and F. sporotrichioides (11 isolates). A set of isolates, which was not determined based on PCR analyses, was whole genome sequenced. Multiple sequence analyses using tef-1α, top1, rpb1, rpb2, tub2, pgk, cam and lsu genes showed that most of them belonged to Equiseti clade. Three cryptic species from this clade: F. clavum, F. flagelliforme and FIESC 31 (lacking Latin binomial) were found on soybean for the first time. This is the first report demonstrating the prevalence of Fusaria on soybean grains in Poland
The Multifaceted Regulation of SnRK2 Kinases
SNF1-related kinases 2 (SnRK2s) are central regulators of plant responses to environmental cues simultaneously playing a pivotal role in the plant development and growth in favorable conditions. They are activated in response to osmotic stress and some of them also to abscisic acid (ABA), the latter being key in ABA signaling. The SnRK2s can be viewed as molecular switches between growth and stress response; therefore, their activity is tightly regulated; needed only for a short time to trigger the response, it has to be induced transiently and otherwise kept at a very low level. This implies a strict and multifaceted control of SnRK2s in plant cells. Despite emerging new information concerning the regulation of SnRK2s, especially those involved in ABA signaling, a lot remains to be uncovered, the regulation of SnRK2s in an ABA-independent manner being particularly understudied. Here, we present an overview of available data, discuss some controversial issues, and provide our perspective on SnRK2 regulation