ABSTRACT
Endocytosis is essential to all eukaryotes, but how cargoes are selected for internalization remains poorly characterized. Extracellular cargoes are thought to be selected by transmembrane receptors that bind intracellular adaptors proteins to initiate endocytosis. Here, we report a mechanism for clathrin-mediated endocytosis (CME) of extracellular lanthanum [La(III)] cargoes, which requires extracellular arabinogalactan proteins (AGPs) that are anchored on the outer face of the plasma membrane. AGPs were colocalized with La(III) on the cell surface and in La(III)-induced endocytic vesicles in Arabidopsis leaf cells. Superresolution imaging showed that La(III) triggered AGP movement across the plasma membrane. AGPs were then colocalized and physically associated with the µ subunit of the intracellular adaptor protein 2 (AP2) complexes. The AGP-AP2 interaction was independent of CME, whereas AGP's internalization required CME and AP2. Moreover, we show that AGP-dependent endocytosis in the presence of La(III) also occurred in human cells. These findings indicate that extracellular AGPs act as conserved CME cargo receptors, thus challenging the current paradigm about endocytosis of extracellular cargoes.
Subject(s)
Endocytosis/genetics , Galactans/metabolism , Lanthanum/pharmacology , Metals, Rare Earth/pharmacology , Adaptor Proteins, Signal Transducing/drug effects , Adaptor Proteins, Signal Transducing/genetics , Cell Membrane/drug effects , Clathrin/chemistry , Endocytosis/drug effects , Galactans/genetics , Humans , Lanthanum/chemistry , Lanthanum/metabolism , Metals, Rare Earth/chemistry , Metals, Rare Earth/metabolism , Multiprotein Complexes/genetics , Multiprotein Complexes/metabolismABSTRACT
With increasing application of rare earth elements (REEs), the resulting environmental safety has attracted extensive attention. When REEs act on plant leaves, REEs can initiate endocytosis in leaf cells, causing more REEs enter plant cells and then severe damage to plants. But when REEs directly act on plant roots, whether and how REEs affect the endocytosis in root cells remain unknown. Here, we characterized effects of lanthanum [La(III)], a REE with high accumulation in environment, on the endocytosis in root cells of Arabidopsis thaliana, and revealed effect mechanism from the perspective of DNA methylation. We found that La(III) enhanced the endocytosis in root cells and the extent of enhancement depended on the dose and time of La(III) exposure: 160⯵Mâ¯>â¯80⯵M >30⯵M (12â¯h); 80⯵Mâ¯>â¯30⯵M >160⯵M (24â¯h); 24â¯hâ¯â¯>â¯12â¯h. La(III)-enhanced endocytosis in root cells resulted from DNA methylation, which was closely related to the expression level of genes encoding DNA methylases/demethylases: CMT3, DRM2 and DNMT2 for 12â¯h, MET1, CMT1, CMT2, CMT3, DRM2, DNMT2, ROS1, DME, DML2, DML5a, and DML5b for 24â¯h. Conversely, enhanced endocytosis also promoted the expression level of genes encoding DNA methylases/demethylases. Our findings provide references for understanding the mechanisms by which REEs impact plants.
Subject(s)
Arabidopsis/metabolism , DNA Methylation/drug effects , Endocytosis/drug effects , Metals, Rare Earth/pharmacology , Plant Roots/metabolism , Arabidopsis Proteins/metabolism , DNA Glycosylases , DNA Methylation/genetics , Gene Expression Regulation, Plant , Lanthanum/metabolism , Lanthanum/pharmacology , Metals, Rare Earth/metabolism , Plant Roots/physiologyABSTRACT
The natural city, which is essential to understand urban physical scale and identify urban sprawling in urban studies, represents the urban functional boundaries of the city defined by human activities rather than the administrative boundaries. Most studies tend to utilize physical environment data such as street networks and remote sensing data to delimitate the natural city, however, such data may not match the real distribution of human activity density in the new cities or even ghost cities in China. This paper suggests aggregating the natural city boundary from the service area polygons of points of interest based on Reilly's Law of Retail Gravitation and the maximum entropy method, since most points of interests provide services for surrounding communities, reflecting the vitality in a bottom-up way. The results indicate that the natural city defined by points of interests shows a high resolution and accuracy, providing a method to define the natural city with POIs.
ABSTRACT
Rare earth element (REE) pollution and acid rain are major global environmental concerns, and their spatial distributions overlap. Thus, both forms of pollution combine to act on plants. Nitrogen is important for plant growth, and nitrate reductase (NR) is a key plant enzyme that catalyzes nitrogen assimilation. Studying the combined effects of REEs and acid rain on plant nitrogen-based nutrients has important environmental significance. Here, soybean (Glycine max) plants, commonly used for toxicological studies, were exposed to lanthanum (La), a REE, and acid rain to study the NR activities and NR transcriptional levels in the roots. To explain how the pollution affected the NR transcriptional level, we simultaneously observed the contents of intracellular La and nutrient elements, protoplast morphology, membrane lipid peroxidation and intracellular pH. A combined treatment of 0.08mmol/L La and pH 4.5 acid rain increased the NR activity, decreased the NR transcriptional level, increased the intracellular nutrient elements' contents and caused deformations in membrane structures. Other combined treatments significantly decreased the aforementioned parameters and caused serious damage to the membrane structures. The variation in the amplitudes of combined treatments was greater than those of individual treatments. Compared with the control and individual treatments, combined treatments increased membrane permeability, the malondialdehyde content, and intracellular H+ and La contents, and with an increasing La concentration or acid strength, the change in amplitude increased. Thus, the combined effects on NR gene transcription in soybean seedling roots were related to the intracellular nutrient elements' contents, protoplast morphology, membranous lipid peroxidation, intracellular pH and La content.
Subject(s)
Acid Rain/adverse effects , Cell Membrane/metabolism , Glycine max/enzymology , Lanthanum/pharmacology , Nitrate Reductase/metabolism , Plant Roots/enzymology , Hydrogen-Ion Concentration , Lanthanum/metabolism , Lipid Peroxidation , Malondialdehyde/metabolism , Membrane Lipids/metabolism , Nitrate Reductase/genetics , Permeability , Plant Roots/drug effects , Plant Roots/metabolism , Protoplasts/drug effects , Protoplasts/pathology , Seedlings/drug effects , Seedlings/enzymology , Glycine max/drug effects , Glycine max/metabolism , Transcription, Genetic/drug effectsABSTRACT
Bisphenol A (BPA), an environmental endocrine disruptor, is an important industrial raw material. The wide use of BPA has increased the risk of BPA release into the environment, and it has become a new environmental pollutant. In this work, the ecological deleterious effects of this new pollutant on soybean roots at different growth stages were investigated by determining the contents of mineral elements (P, K, Ca, and Mg) and analyzing root activity and the activities of critical respiratory enzymes (hexokinase, phosphofructokinase, pyruvate kinase, and isocitrate dehydrogenase). Our results revealed that low dose (1.5mg/L) of BPA increased the levels of P, K, Mg, and Ca in soybean roots at different growth stages. Whereas, high doses (6.0 and 12.0mg/L) of BPA decreased the levels of P, K, and Mg contents in a dose-dependent manner. BPA had a promotive effect on the content of Ca in soybean roots. Synchronous observation showed that the aforementioned dual response to BPA were also observed in the root activity and respiratory enzyme activities. The effects of BPA on the mineral element contents, root activity and respiratory enzyme activities in soybean roots at different growth stages followed the order: flowering and podding stage>seed-filling stage>seedling stage (mineral element contents); seedling stage>flowering and podding stage>seed-filling stage (root activity and respiratory enzyme activities). In a word, the response of plant root activity and respiratory enzyme activities to BPA pollution is a pathway of BPA affecting mineral element contents in plant roots.